Indolizine compounds, a process for their preparation and pharmaceutical compositions containing them

ABSTRACT

Compounds of formula (I) wherein Ra, Rb, Rc, Rd, T, R3, R4, R5, X, Y and Het are as defined in the description. 
     
       
         
         
             
             
         
       
     
     Medicinal products containing the same which are useful in treating pathologies involving a deficit in apoptosis, such as cancer, auto-immune diseases, and diseases of the immune system.

The present invention relates to new indolizine compounds, to a processfor their preparation and to pharmaceutical compositions containingthem.

The compounds of the present invention are new and have very valuablepharmacological characteristics in the field of apoptosis andcancerology.

Apoptosis, or programmed cell death, is a physiological process that iscrucial for embryonic development and maintenance of tissue homeostasis.

Apoptotic-type cell death involves morphological changes such ascondensation of the nucleus, DNA fragmentation and also biochemicalphenomena such as the activation of caspases which cause damage to keystructural components of the cell, so inducing its disassembly anddeath. Regulation of the process of apoptosis is complex and involvesthe activation or repression of several intracellular signallingpathways (Cory S. et al., Nature Review Cancer, 2002, 2, 647-656).

Deregulation of apoptosis is involved in certain pathologies. Increasedapoptosis is associated with neurodegenerative diseases such asParkinson's disease, Alzheimer's disease and ischaemia. Conversely,deficits in the implementation of apoptosis play a significant role inthe development of cancers and their chemoresistance, in auto-immunediseases, inflammatory diseases and viral infections. Accordingly,absence of apoptosis is one of the phenotypic signatures of cancer(Hanahan D. et al., Cell 2000, 100, 57-70).

The anti-apoptotic proteins of the Bcl-2 family are associated withnumerous pathologies. The involvement of proteins of the Bcl-2 family isdescribed in numerous types of cancer, such as colorectal cancer, breastcancer, small-cell lung cancer, non-small-cell lung cancer, bladdercancer, ovarian cancer, prostate cancer, chronic lymphoid leukaemia,follicular lymphoma, myeloma, prostate cancer, etc. Overexpression ofthe anti-apoptotic proteins of the Bcl-2 family is involved intumorigenesis, in resistance to chemotherapy and in the clinicalprognosis of patients affected by cancer. There is, therefore, atherapeutic need for compounds that inhibit the anti-apoptotic activityof the proteins of the Bcl-2 family.

In addition to being new, the compounds of the present invention havepro-apoptotic properties making it possible to use them in pathologiesinvolving a defect in apoptosis, such as, for example, in the treatmentof cancer, auto-immune diseases and diseases of the immune system.

The present invention relates more especially to compounds of formula(I):

wherein:

-   -   X and Y represent a carbon atom or a nitrogen atom, it being        understood that they may not simultaneously represent two        carbons atoms or two nitrogen atoms,    -   the Het moiety of the group

-   -    represents an optionally substituted, aromatic or non-aromatic        ring composed of 5, 6 or 7 ring members, which may contain, in        addition to the nitrogen represented by X or by Y, from one to 3        hetero atoms selected independently from oxygen, sulphur and        nitrogen, it being understood that the nitrogen in question may        be substituted by a group representing a hydrogen atom, a linear        or branched (C₁-C₆)alkyl group or a group —C(O)—O-Alk wherein        Alk is a linear or branched (C₁-C₆)alkyl group,    -   T represents a hydrogen atom, a linear or branched (C₁-C₆)alkyl        group optionally substituted by from one to three halogen atoms,        a group (C₂-C₄)alkyl-NR₁R₂, or a group (C₁-C₄)alkyl-OR₆,    -   R₁ and R₂ independently of one another represent a hydrogen atom        or a linear or branched (C₁-C₆)alkyl group, or R₁ and R₂ form        with the nitrogen atom carrying them a heterocycloalkyl,    -   R₃ represents a linear or branched (C₁-C₆)alkyl group, a linear        or branched (C₂-C₆)alkenyl group, a linear or branched        (C₂-C₆)alkynyl group, a cycloalkyl group, a        (C₃-C₁₀)cycloalkyl-(C₁-C₆)alkyl group wherein the alkyl moiety        is linear or branched, a heterocycloalkyl group, an aryl group        or a heteroaryl group, it being understood that one or more of        the carbon atoms of the preceding groups, or of their possible        substituents, may be deuterated,    -   R₄ represents an aryl group, a heteroaryl group, a cycloalkyl        group or a linear or branched (C₁-C₆)alkyl group, it being        understood that one or more of the carbon atoms of the preceding        groups, or of their possible substituents, may be deuterated,    -   R₅ represents a hydrogen or halogen atom, a linear or branched        (C₁-C₆)alkyl group, or a linear or branched (C₁-C₆)alkoxy group,    -   R₆ represents a hydrogen atom or a linear or branched        (C₁-C₆)alkyl group,    -   R_(a), R_(b), R_(c) and R_(d), each independently of the others,        represent R₇, a halogen atom, a linear or branched (C₁-C₆)alkoxy        group, a hydroxy group, a linear or branched        (C₁-C₆)polyhaloalkyl group, a trifluoromethoxy group, —NR₇R₇′,        nitro, R₇—CO—(C₀-C₆)alkyl-, R₇—CO—NH—(C₀-C₆)alkyl-,        NR₇R₇′—CO—(C₀-C₆)alkyl-, NR₇R₇′—CO—(C₀-C₆)alkyl-O—,        R₇—SO₂—NH—(C₀-C₆)alkyl-, R₇—NH—CO—NH—(C₀-C₆)alkyl-,        R₇—O—CO—NH—(C₀-C₆)alkyl-, a heterocycloalkyl group, or the        substituents of one of the pairs (R_(a),R_(b)), (R_(b),R_(c)) or        (R_(c),R_(d)) form together with the carbon atoms carrying them        a ring composed of from 5 to 7 ring members, which may contain        from one to 2 hetero atoms selected from oxygen and sulphur, it        also being understood that one or more carbon atoms of the ring        defined hereinbefore may be deuterated or substituted by from        one to 3 groups selected from halogen and linear or branched        (C₁-C₆)alkyl,    -   R₇ and R₇′, each independently of the other, represent a        hydrogen, a linear or branched (C₁-C₆)alkyl, a linear or        branched (C₂-C₆)alkenyl, a linear or branched (C₂-C₆)alkynyl, an        aryl or a heteroaryl, or R₇ and R₇′, together with the nitrogen        atom carrying them, form a heterocycle composed of from 5 to 7        ring members,        it being understood that:    -   “aryl” means a phenyl, naphthyl, biphenyl or indenyl group,    -   “heteroaryl” means any mono- or bi-cyclic group composed of from        5 to 10 ring members, having at least one aromatic moiety and        containing from 1 to 4 hetero atoms selected from oxygen,        sulphur and nitrogen (including quaternary nitrogens),    -   “cycloalkyl” means any mono- or bi-cyclic, non-aromatic,        carbocyclic group containing from 3 to 10 ring members,    -   “heterocycloalkyl” means any mono- or bi-cyclic, non-aromatic,        condensed or spiro group containing from 3 to 10 ring members        and containing from 1 to 3 hetero atoms selected from oxygen,        sulphur, SO, SO₂ and nitrogen,        it being possible for the aryl, heteroaryl, cycloalkyl and        heterocycloalkyl groups so defined and the groups alkyl,        alkenyl, alkynyl and alkoxy to be substituted by from 1 to 3        groups selected from optionally substituted, linear or branched        (C₁-C₆)alkyl, (C₃-C₆)spiro, optionally substituted, linear or        branched (C₁-C₆)alkoxy, (C₁-C₆)alkyl-S—, hydroxy, oxo (or        N-oxide where appropriate), nitro, cyano, —COOR′, —OCOR′, NR′R″,        linear or branched (C₁-C₆)polyhaloalkyl, trifluoromethoxy,        (C₁-C₆)alkylsulphonyl, halogen, optionally substituted aryl,        heteroaryl, aryloxy, arylthio, cycloalkyl, heterocycloalkyl        optionally substituted by one or more halogen atoms or alkyl        groups, it being understood that R′ and R″, each independently        of the other, represent a hydrogen atom or an optionally        substituted, linear or branched (C₁-C₆)alkyl group,        it being possible for the Het moiety of the group

Het defined in formula (I) to be substituted by from one to three groupsselected from linear or branched (C₁-C₆)alkyl, hydroxy, linear orbranched (C₁-C₆)alkoxy, NR₁′R₁″ and halogen, it being understood thatR₁′ and R₁″ are as defined for the groups R′ and R″ mentionedhereinbefore,to their enantiomers and diastereoisomers, and to addition salts thereofwith a pharmaceutically acceptable acid or base.

Among the pharmaceutically acceptable acids there may be mentioned,without implying any limitation, hydrochloric acid, hydrobromic acid,sulphuric acid, phosphonic acid, acetic acid, trifluoroacetic acid,lactic acid, pyruvic acid, malonic acid, succinic acid, glutaric acid,fumaric acid, tartaric acid, maleic acid, citric acid, ascorbic acid,oxalic acid, methanesulphonic acid, camphoric acid etc.

Among the pharmaceutically acceptable bases there may be mentioned,without implying any limitation, sodium hydroxide, potassium hydroxide,triethylamine, tert-butylamine etc.

Advantageously, the group:

represents one of the following groups: 5,6,7,8-tetrahydroindolizineoptionally substituted by an amino group; indolizine;1,2,3,4-tetrahydropyrrolo[1,2-a]pyrazine optionally substituted by amethyl; pyrrolo[1,2-a]pyrimidine. The groups5,6,7,8-tetrahydroindolizine and indolizine are more especiallypreferred.

In preferred compounds of the invention, T represents a hydrogen atom, amethyl group (and more especially an (R)-methyl), a group2-(morpholin-4-yl)ethyl, 3-(morpholin-4-yl)propyl, —CH₂—OH,2-aminoethyl, 2-(3,3-difluoropiperidin-1-yl)ethyl,2-[(2,2-difluoroethyl)amino]ethyl or 2-(3-methoxyazetidin-1-yl)ethyl.

Preferably, R_(a) and R_(d) each represent a hydrogen atom and(R_(b),R_(c)), together with the carbon atoms carrying them, form a1,3-dioxolane group or a 1,4-dioxane group; or R_(a), R_(c) and R_(d)each represent a hydrogen atom and R_(b) represents a hydrogen, ahalogen, a methyl or a methoxy; or R_(a), R_(b) and R_(d) each representa hydrogen atom and R_(c) represents a hydroxy or methoxy group. Evenmore preferably, R_(a) and R_(d) each represent a hydrogen atom and(R_(b),R_(c)), together with the carbon atoms carrying them, form a1,3-dioxolane group; or R_(a), R_(c) and R_(d) each represent a hydrogenatom and R_(b) represents a halogen. Preference is given to the R₄ groupbeing a 4-hydroxyphenyl.

In preferred compounds of the invention, R₃ represents a linear(C₁-C₆)alkyl, aryl or heteroaryl group, it being possible for the lattertwo groups to be substituted by from one to three groups selected fromhalogen, linear or branched (C₁-C₆)alkyl, linear or branched(C₁-C₆)alkoxy, cyano and heterocycloalkyl-(C₁-C₆)alkyl wherein the alkylmoiety is linear or branched. Even more preferably, R₃ represents aheteroaryl group selected from the following group: 1H-indole,2,3-dihydro-1H-indole, 1H-indazole, pyridine, 1H-pyrrolo[2,3-b]pyridine,1H-pyrazole, imidazo[1,2-a]pyridine, pyrazolo[1,5-a]pyrimidine,[1,2,4]triazolo[1,5-a]pyrimidine, and 1H-pyrazolo[3,4-b]pyridine, all ofwhich may be substituted by a linear or branched (C₁-C₆)alkyl group.

Preferred compounds according to the invention are included in thefollowing group:

-   N-(4-hydroxyphenyl)-3-(6-{[(3R)-3-methyl-3,4-dihydroisoquinolin-2(1H)-yl]carbonyl}-1,3-benzodioxol-5-yl)-N-{1-[2-(morpholin-4-yl)ethyl]-1H-indol-5-yl}-5,6,7,8-tetrahydroindolizine-1-carboxamide,-   N-(4-hydroxyphenyl)-3-(6-{[(3S)-3-[2-(morpholin-4-yl)ethyl]-3,4-dihydroisoquinolin-2(1H)-yl]carbonyl}-1,3-benzodioxol-5-yl)-N-phenyl-5,6,7,8-tetrahydroindolizine-1-carboxamide,-   N-{3-fluoro-4-[2-(morpholin-4-yl)ethoxy]phenyl}-N-(4-hydroxyphenyl)-3-(6-{[(3R)-3-methyl-3,4-dihydroisoquinolin-2(1H)-yl]carbonyl}-1,3-benzodioxol-5-yl)indolizine-1-carboxamide,-   N-(4-hydroxyphenyl)-3-(6-{[(3R)-3-methyl-3,4-dihydroisoquinolin-2(1H)-yl]carbonyl}-1,3-benzodioxol-5-yl)-N-(pyridin-4-yl)indolizine-1-carboxamide,-   N-(4-hydroxyphenyl)-3-(6-{[(3R)-3-methyl-3,4-dihydroisoquinolin-2(1H)-yl]carbonyl}-1,3-benzodioxol-5-yl)-N-(2-methylpyridin-4-yl)indolizine-1-carboxamide,-   N-(4-hydroxyphenyl)-3-(6-{[(3R)-3-methyl-3,4-dihydroisoquinolin-2(1H)-yl]carbonyl}-1,3-benzodioxol-5-yl)-N-(1-methyl-1H-pyrrolo[2,3-b]pyridin-5-yl)indolizine-1-carboxamide,-   N-(4-hydroxyphenyl)-3-(6-{[(3R)-3-[3-(morpholin-4-yl)propyl]-3,4-dihydroisoquinolin-2(1H)-yl]carbonyl}-1,3-benzodioxol-5-yl)-N-phenyl-5,6,7,8-tetrahydroindolizine-1-carboxamide,-   N-(2,6-dimethylpyridin-4-yl)-N-(4-hydroxyphenyl)-3-(6-{[(3R)-3-methyl-3,4-dihydroisoquinolin-2(1H)-yl]carbonyl}-1,3-benzodioxol-5-yl)indolizine-1-carboxamide,-   N-(4-hydroxyphenyl)-3-(6-{[(3R)-3-methyl-3,4-dihydroisoquinolin-2(1H)-yl]carbonyl}-1,3-benzodioxol-5-yl)-N-(pyridin-4-yl)-5,6,7,8-tetrahydroindolizine-1-carboxamide,-   3-(5-chloro-2-{[(3R)-3-methyl-3,4-dihydroisoquinolin-2(1H)-yl]carbonyl}phenyl)-N-(4-hydroxyphenyl)-N-(1-methyl-1H-pyrrolo[2,3-b]pyridin-5-yl)indolizine-1-carboxamide,-   N-(4-hydroxyphenyl)-N-(2-methoxypyridin-4-yl)-3-(6-{[(3R)-3-methyl-3,4-dihydroisoquinolin-2(1H)-yl]carbonyl}-1,3-benzodioxol-5-yl)indolizine-1-carboxamide,    their enantiomers and diastereoisomers, and addition salts thereof    with a pharmaceutically acceptable acid or base.

The invention relates also to a process for the preparation of compoundsof formula (I), which process is characterised in that there is used asstarting material the compound of formula (II):

wherein R_(a), R_(b), R_(c) and R_(d) are as defined for formula (I),which compound of formula (II) is subjected to a Heck reaction, in anaqueous or organic medium, in the presence of a palladium catalyst, of abase, of a phosphine and of the compound of formula (III):

wherein the groups X, Y and Het are as defined for formula (I),to obtain the compound of formula (IV):

wherein R_(a), R_(b), R_(c), R_(d), X, Y and Het are as defined forformula (I),the aldehyde function of which compound of formula (IV) is oxidised to acarboxylic acid to form the compound of formula (V):

wherein R_(a), R_(b), R_(c), R_(d), X, Y and Het are as defined forformula (I),which compound of formula (V) is then subjected to peptide coupling witha compound of formula (VI):

wherein T and R₅ are as defined for formula (I),to yield the compound of formula (VII):

wherein R_(a), R_(b), R_(c), R_(d), T, R₅, X, Y and Het are as definedfor formula (I),the ester function of which compound of formula (VII) is hydrolysed toyield the corresponding carboxylic acid or carboxylate, which may beconverted into an acid derivative such as the corresponding acylchloride or anhydride before being coupled with an amine NHR₃R₄ whereinR₃ and R₄ have the same meanings as for formula (I), to yield thecompound of formula (I),which compound of formula (I) may be purified according to aconventional separation technique, which is converted, if desired, intoits addition salts with a pharmaceutically acceptable acid or base andwhich is optionally separated into its isomers according to aconventional separation technique,it being understood that, at any time considered appropriate in thecourse of the above-described process, certain groups (hydroxy, amino .. . ) of the reagents or intermediates of synthesis may be protected andthen deprotected according to the requirements of synthesis.

More particularly, when one of the groups R₃ or R₄ of the amine NHR₃R₄is substituted by a hydroxy function, the latter may be subjectedbeforehand to a protection reaction prior to any coupling with thecarboxylic acid formed from the compound of formula (VII), or with acorresponding acid derivative thereof, the resulting protected compoundof formula (I) subsequently undergoes a deprotection reaction and isthen optionally converted into one of its addition salts with apharmaceutically acceptable acid or base.

The compounds of formulae (II), (III), (VI) and the amine NHR₃R₄ areeither commercially available or can be obtained by the person skilledin the art using conventional chemical reactions described in theliterature.

Pharmacological study of the compounds of the invention has shown thatthey have pro-apoptotic properties. The ability to reactivate theapoptotic process in cancerous cells is of major therapeutic interest inthe treatment of cancers, auto-immune diseases and diseases of theimmune system.

More especially, the compounds according to the invention will be usefulin the treatment of chemo- or radio-resistant cancers, and in malignanthaemopathies and small-cell lung cancer.

Among the cancer treatments envisaged there may be mentioned, withoutimplying any limitation, cancers of the bladder, brain, breast anduterus, chronic lymphoid leukaemias, colorectal cancer, cancers of theœsophagus and liver, lymphoblastic leukaemias, non-Hodgkin lymphomas,melanomas, malignant haemopathies, myelomas, ovarian cancer,non-small-cell lung cancer, prostate cancer and small-cell lung cancer.Among non-Hodgkin lymphomas, there may be mentioned more preferablyfollicular lymphomas, mantle cell lymphomas, diffuse large B-celllymphomas, small lymphocytic lymphomas and marginal zone B-celllymphomas.

The present invention relates also to pharmaceutical compositionscomprising at least one compound of formula (I) in combination with oneor more pharmaceutically acceptable excipients.

Among the pharmaceutical compositions according to the invention theremay be mentioned more especially those that are suitable for oral,parenteral, nasal, per- or trans-cutaneous, rectal, perlingual, ocularor respiratory administration, especially tablets or dragées, sublingualtablets, sachets, paquets, capsules, glossettes, lozenges,suppositories, creams, ointments, dermal gels, and drinkable orinjectable ampoules.

The dosage varies according to the sex, age and weight of the patient,the administration route, the nature of the therapeutic indication, orof any associated treatments, and ranges from 0.01 mg to 1 g per 24hours in one or more administrations.

Furthermore, the present invention relates also to the association of acompound of formula (I) with an anticancer agent selected from genotoxicagents, mitotic poisons, anti-metabolites, proteasome inhibitors, kinaseinhibitors and antibodies, and also to pharmaceutical compositionscomprising that type of association and their use in the manufacture ofmedicaments for use in the treatment of cancer.

The compounds of the invention may also be used in association withradiotherapy in the treatment of cancer.

Finally, the compounds of the invention may be linked to monoclonalantibodies or fragments thereof or linked to scaffold proteins that canbe related or not to monoclonal antibodies.

Antibody fragments must be understood as fragments of Fv, scFv, Fab,F(ab′)2, F(ab′), scFv-Fc type or diabodies, which generally have thesame specificity of binding as the antibody from which they aredescended. According to the present invention, antibody fragments of theinvention can be obtained starting from antibodies by methods such asdigestion by enzymes, such as pepsin or papain, and/or by cleavage ofthe disulfide bridges by chemical reduction. In another manner, theantibody fragments comprised in the present invention can be obtained bytechniques of genetic recombination likewise well known to the personskilled in the art or else by peptide synthesis by means of, forexample, automatic peptide synthesizers such as those supplied by thecompany Applied Biosystems, etc.

Scaffold proteins that can be related or not to monoclonal antibodiesare understood to mean a protein that contains or not an immunoglobulinfold and that yields a binding capacity similar to a monoclonalantibody. The man skilled in the art knows how to select the proteinscaffold. More particularly, it is known that, to be selected, such ascaffold should display several features as follow (Skerra A., J. Mol.Recogn., 13, 2000, 167-187): phylogenetically good conservation, robustarchitecture with a well-known three-dimensional molecular organization(such as, for example, crystallography or NMR), small size, no or only alow degree of post-translational modifications, easy to produce, expressand purify. Such a protein scaffold can be, but without limitation, astructure selected from the group consisting in fibronectin andpreferentially the tenth fibronectin type III domain (FNfn10),lipocalin, anticalin (Skerra A., J. Biotechnol., 2001, 74(4):257-75),the protein Z derivative from the domain B of staphylococcal protein A,thioredoxin A or any protein with a repeated domain such as an “ankyrinrepeat” (Kohl et al., PNAS, 2003, vol. 100, No. 4, 1700-1705),“armadillo repeat”, “leucine-rich repeat” or “tetratricopeptide repeat”.

There could also be mentioned a scaffold derivative from toxins (suchas, for example, scorpion, insect, plant or mollusc toxins) or proteininhibitors of neuronal nitric oxide synthase (PIN).

The following Preparations and Examples illustrate the invention withoutlimiting it in any way.

General Procedures

All reagents and anhydrous solvents are obtained from commercial sourcesand were used without further purification or drying. Flashchromatography is performed on an ISCO CombiFlash Rf 200i apparatus withpre-packed silica-gel cartridges (SiliaSep™ F60 (40-63 m, 60 Å). Thinlayer chromatography was conducted with 5×10 cm plates coated with MerckType 60 F₂₅₄ silica gel. Microwave heating was performed with a CEMDiscover® SP apparatus.

Analytical LC-MS

The compounds of the invention were characterized by high-performanceliquid chromatography coupled with mass spectroscopy (HPLC-MS) on eitheran Agilent HP1200 rapid-resolution apparatus coupled to a 6140 massdetector with a multi-mode source (m/z range 150 to 1000 atomic massunits or amu) or an Agilent HP1100 apparatus coupled to a 1946D massdetector with an electrospray ionisation source (m/z range 150 to 1000amu). The conditions and methods listed below are identical for bothmachines.

-   Detection: UV detection at 230, 254 and 270 nm.-   Injection Volume: 2 μL-   Mobile Phases: A—Water+10 mMol/ammonium formate+0.08% (v/v) formic    acid at pH ca 3.5.    -   B—95% Acetonitrile+5% A+0.08% (v/v) formic acid

Method A (3.75 min; either positive (pos) or positive and negative(pos/neg) ionisation)

-   Column: Gemini 5 m, C18, 30 mm×4.6 mm (Phenomenex).-   Temperature: 35° C.

Gradient:

Flow Time (min) Solvent A (%) Solvent B (%) (mL/min) 0 95 5 2 0.25 95 52 2.50 95 5 2 2.55 5 95 3 3.60 5 95 3 3.65 5 95 2 3.70 95 5 2 3.75 95 52

Method B (1.9 min; either positive (pos) or positive and negative(pos/neg) ionisation)

-   Column: Gemini 5 μm, C18, 30 mm×4.6 mm (Phenomenex).-   Temperature: 35° C.

Gradient:

Flow Time (min) Solvent A (%) Solvent B (%) (mL/min) 0 95 5 1.1 0.12 955 1.1 1.30 5 95 1.1 1.35 5 95 1.7 1.85 5 95 1.7 1.90 5 95 1.1 1.95 95 51.1

Preparation 1:6-[1-(Methoxycarbonyl)-5,6,7,8-tetrahydro-3-indolizinyl]-1,3-benzodioxole-5-carboxylicacid Step A: 1-Formyl-2-piperidine-carboxylic acid

To a solution of 40 g of a racemic mixture of 2-piperidine-carboxylicacid (0.310 mmol) in 300 mL of formic acid placed at 0° C. there areadded, dropwise, 200 mL (2.15 mmol) of acetic anhydride. The batch isthen stirred at ambient temperature overnight. Then, the reactionmixture is cooled to 0° C., hydrolysed by adding 250 mL of water, andstirred for half an hour at 0° C. before being concentrated to dryness.The oil thereby obtained is taken up in 200 mL of methanol and thenconcentrated to dryness. The title product is obtained in the form of anoil in a yield of 98%. It is used directly, without being otherwisepurified, in the next Step.

¹H NMR: δ (400 MHz; dmso-d6; 300K): 13.0 (m, 1H OH); 8.0-8.05 (2s, 1Haldehyde); 4.9-4.5 (2d, 1H α to the N and COOH); 4.1-2.6 (m, 2H α to theN); 2.2-1.2 (m, 6H piperidine)

IR: ν: —OH: 2000-3000 cm⁻¹ acid; ν: >C═O 1703 cm⁻¹ wide band

Step B: Methyl 5,6,7,8-tetrahydro-1-indolizine-carboxylate

To a solution of 10 g of the carboxylic acid obtained in Step A (63.6mmol) in 65 mL of dichloromethane there are successively added 13.4 g oftosyl chloride (70.4 mmol), 11.5 mL of methyl 2-chloroacrylate (113.5mmol) and then, dropwise, 17.8 mL of N,N,N-triethylamine (127.2 mmol).The reaction mixture is then refluxed for 1 hour 30 minutes. It is thenplaced at ambient temperature, and there are then added 5 mL of methyl2-chloroacrylate (48.9 mmol) and, dropwise, 9 mL of N,N,N-triethylamine(64 mmol). The batch is refluxed overnight.

The reaction mixture is then diluted with methylene chloride, washedsuccessively with 1N HCl solution, saturated NaHCO₃ solution and thensaturated NaCl solution until a neutral pH is obtained. The organicphase is then dried over MgSO₄, filtered, concentrated to dryness andpurified by chromatography over silica gel (heptane/AcOEt gradient). Thetitle product is obtained in the form of an oil.

¹H NMR: δ (400 MHz; CDCl₃; 300K): 6.55-6.40 (d, 2H,tetrahydroindolizine); 3.91 (t, 3H methyl ester); 3.78 (s, 3Htetrahydroindolizine); 3.08 (t, 2H, tetrahydroindolizine); 1.95-1.85 (m,4H, tetrahydroindolizine)

IR: ν:>C═O 1692 cm⁻¹ ester

Step C: Methyl3-(6-formyl-1,3-benzodioxol-5-yl)-5,6,7,8-tetrahydro-1-indolizine-carboxylate

To a solution of 6.4 g of the ester obtained in Step B (35.7 mmol) in 12mL of N,N-dimethylacetamide, there are successively added 12.3 g of6-bromo-1,3-benzodioxole-5-carbaldehyde (53.6 mmol) and 7 g of potassiumacetate (71.4 mmol), and then the batch is stirred under argon for 20minutes. There are then added 1.3 g of palladium catalystPdCl₂(PPh₃)₂(1.8 mmol). The reaction mixture is then heated at 130° C.for one hour before adding 139 μL of H₂O thereto. Heating is maintainedat that same temperature overnight. The mixture is allowed to return toambient temperature and it is then diluted with AcOEt. Animal charcoal(2 g per g of product) is added and the batch is stirred at ambienttemperature for 1 hour and then filtered. The organic phase is thenwashed with water, dried over magnesium sulphate and concentrated todryness. The crude product thereby obtained is purified over silica gel(heptane/ACOEt gradient). The title product is obtained in the form ofan oil.

¹H NMR: δ:(400 MHz; dmso-d6; 353° K): 9.65 (s, 1H, H aldehyde); 7.3-7.15(2s, 2H, aromatic Hs); 6.45 (s, 1H tetrahydroindolizine); 6.20 (s, 2Hmethylenedioxy); 3.70 (s, 3H methyl ester); 3.5-4.0 (m, 2Htetrahydroindolizine); 3.05 (m, 2H tetrahydroindolizine); 1.85 (m, 4Htetrahydroindolizine)

IR: ν: >C═O 1695 cm⁻¹ ester; ν: >C═O 1674 cm⁻¹

Step D:6-[1-(Methoxycarbonyl)-5,6,7,8-tetrahydro-3-indolizinyl]-1,3-benzodioxole-5-carboxylicacid

A solution containing 3.37 g of the compound obtained in Step C (10.3mmol) in 9.3 mL of acetone and 8.8 mL (80.24 mmol) of 2-methyl-2-buteneis prepared and placed at 0° C. There are added, dropwise, 9.3 mL of anaqueous solution containing a mixture of 3.3 g of NaClO₂ (36.05 mmol)and 3.6 g of Na₂PO₄ (25.75 mmol). The batch is then stirred at ambienttemperature for 7 hours. The reaction mixture is then concentrated inorder to remove the acetone. Then solid then obtained is filtered off,washed with water and then dried at 40° C. in vacuo overnight. The titleproduct is obtained in the form of a solid, which is subsequently usedwithout being otherwise purified.

¹H NMR: δ (400 MHz; dmso-d6; 300K): 12.10 (m, 1H, H carboxylic acid);7.40-6.88 (2s, 2H, aromatic Hs); 6.20 (s, 1H, H tetrahydroindolizine);6.18 (s, 2H, H methylenedioxy); 3.70 (s, 3H, methyl ester); 3.55 (t, 2Htetrahydroindolizine); 3.00 (t, 2H tetrahydroindolizine); 1.80 (m, 4H, Htetrahydroindolizine)

IR: ν: —OH: 3000-2000 cm⁻¹ acid; ν: >C═O 1686-1676 cm⁻¹ ester+acid;ν: >C═C<1608 cm⁻¹

Preparation 2:2-[2-(tert-Butoxycarbonyl)-8-(methoxycarbonyl)-1,2,3,4-tetrahydropyrrolo[1,2-a]pyrazin-6-yl]-4-chlorobenzoicacid Step A: 1-tert-Butyl 3-methyl 4-formyl-1,3-piperazinedicarboxylate

To a solution of pentafluorophenol in 520 mL of anhydrous ether placedat 0° C. there are successively added 49 g of1-ethyl-3-(3′-dimethylaminopropyl)-carbodiimide (286 mmol) in portionsand 12 mL of formic acid (312 mmol). The batch is stirred at ambienttemperature for 2 hours. There is then added a mixture of 32 g of1-tert-butyl 3-methyl 1,3-piperazinedicarboxylate (130 mmol) and 18 mLof triethylamine (130 mmol) dissolved in 520 mL of CH₂Cl₂. The batch isstirred overnight at ambient temperature. The reaction mixture ishydrolysed with 1N aqueous HCl solution and extracted with CH₂Cl₂. Theorganic phases are then combined and then washed with saturated aqueousNaHCO₃ solution and then with saturated aqueous NaCl solution untilneutral. After drying over MgSO₄, filtering and concentrating todryness, the product is isolated by chromatography over silica gel(petroleum ether/AcOEt gradient: 0-30%). The title product is obtainedin the form of an oil.

IR: ν: C═O: 1674-1745 cm⁻¹

m/z (C₁₂H₂₀N₂O₅): 272.1 (M+); 295.121 (M+Na)⁺; 567.253 (2M+Na)⁺

Step B: Lithium 4-(tert-butoxycarbonyl)-1-formyl-2-piperazinecarboxylate

To a solution of 28 g of the compound obtained in Step A (103 mmol) in515 mL of dioxane there are added 4.8 g of LiOH (113 mmol) dissolved in100 mL of H₂O. The batch is stirred at ambient temperature for 4 hours.The reaction mixture is then concentrated to dryness and thenco-evaporated several times with ethyl acetate. The title product isobtained in the form of a solid and is used directly in the followingcyclisation step.

¹³C NMR: δ (500 MHz; dmso-d6; 300K): 46 (s, C piperazine); 42-38 (m, Cpiperazine); 58-53 (s, C piperazine); 28.5 (s, C ^(t)Bu)

IR: ν: C═O: 1650 cm⁻¹; 2800 cm⁻¹

Step C: 2-tert-Butyl 8-methyl3,4-dihydropyrrolo[1,2-a]pyrazine-2,8(1H)-dicarboxylate

To a suspension of 29 g of the compound obtained in Step B (103 mmol) in800 mL of dichloromethane there are successively added 24 g of tosylchloride (124 mmol), 12.6 mL of methyl 2-chloroacrylate (124 mmol) andthen 35 mL of triethylamine (247 mmol). The batch is stirred at refluxfor 2 hours. After cooling, the reaction mixture is diluted with ethylacetate and the organic phase is washed with saturated NaCl solutionuntil neutral. After drying over MgSO₄, filtering and concentrating todryness, the title product is isolated by chromatography over silica gel(petroleum ether/AcOEt gradient: 0-20%) in the form of a solid.

¹H NMR: δ (400 MHz; dmso-d6; 300K): 6.8-6.43 (m, 2H, H pyrrole);4.75-3.75 (m, 6H, H piperazine); 3.73 (s, 3H, H COOCH3); 1.48 (s, 9H, H^(t)Bu)

IR: ν: C═O (conjugated ester): 1712 cm⁻¹; C═O (carbamate): 1677 cm⁻¹

Step D:2-[2-(tert-Butoxycarbonyl)-8-(methoxycarbonyl)-1,2,3,4-tetrahydropyrrolo[1,2-a]pyrazin-6-yl]-4-chlorobenzoicacid

The procedure is in accordance with the protocol described in Steps Cand D of Preparation 1, replacing the6-bromo-1,3-benzodioxole-5-carbaldehyde used in Step C with2-bromo-4-chlorobenzaldehyde.

Preparation 3:4-Chloro-2-[1-(methoxycarbonyl)-5,6,7,8-tetrahydro-3-indolizinyl]-benzoicacid

The procedure is in accordance with the process of Preparation 1,replacing the 6-bromo-1,3-benzodioxole-5-carbaldehyde used in Step Cwith 2-bromo-4-chlorobenzaldehyde.

Preparation 4:7-[2-(tert-Butoxycarbonyl)-8-(methoxycarbonyl)-1,2,3,4-tetrahydropyrrolo[1,2-a]pyrazin-6-yl]-2,3-dihydro-1,4-benzodioxine-6-carboxylicacid Step A: 2-tert-Butyl 8-methyl3,4-dihydropyrrolo[1,2-a]pyrazine-2,8(1H)-dicarboxylate

The procedure is in accordance with the process described in Steps A-Cof Preparation 2.

¹H NMR: δ (400 MHz; dmso-d6; 300K): 6.8-6.43 (m, 2H, H pyrrole);4.75-3.75 (m, 6H, H piperazine); 3.73 (s, 3H, H COOCH3); 1.48 (s, 9H, H^(t)Bu)

IR: ν: C═O (conjugated ester): 1712 cm⁻¹; C═O (carbamate): 1677 cm⁻¹

Step B:7-[2-(tert-Butoxycarbonyl)-8-(methoxycarbonyl)-1,2,3,4-tetrahydropyrrolo[1,2-a]pyrazin-6-yl]-2,3-dihydro-1,4-benzodioxine-6-carboxylicacid

The procedure is in accordance with the protocol described in Steps Cand D of Preparation 1, replacing the6-bromo-1,3-benzodioxole-5-carbaldehyde used in Step C with7-bromo-2,3-dihydro-1,4-benzodioxine-6-carbaldehyde.

Preparation 5: 4-Chloro-2-[1-(methoxycarbonyl)-3-indolizinyl]benzoicacid Step A: 1-(Carboxymethyl)-1,2-dihydropyridinium bromide

To a solution of 16.2 mL of pyridine (200 mmol) in 120 mL of ethylacetate there are added, in portions, 27.8 g (200 mmol) of bromoaceticacid. The batch is then stirred at ambient temperature overnight. Theprecipitate thereby obtained is filtered off and then washed with coldethyl acetate. After drying, the title product is obtained in the formof a powder which is used directly in the next Step.

¹H NMR: δ (400 MHz; dmso-d6; 300K): 9.15 (d, 2H, aromatic Hs pyridine);8.7 (t, 1H, aromatic H); 8.25 (t, 2H, aromatic H); 5.65 (s, 2H, HCH₂COOH)

IR: ν: C═O: 1732 cm⁻¹; —OH acid: 2800 cm⁻¹

Step B: Methyl 1-indolizinecarboxylate

To a suspension of 6.55 g of the pyridinium salt obtained in Step A (30mmol) in 240 mL of toluene there are successively added 16.7 mL ofmethyl acrylate (150 mmol), 4.2 mL of triethylamine (30 mmol) and then,in portions, 20.9 g of MnO₂ (240 mmol). The batch is then heated at 90°C. for 3 hours. After cooling, the reaction mixture is filtered over acake of Celite and concentrated to dryness. The title product is thenisolated by purification over silica gel (heptane/AcOEt gradient: 0-10%)in the form of an oil which crystallises in the cold state.

¹H NMR: δ (300 MHz; dmso-d6; 300K): 8.5 (d, 1H, H indolizine); 8.05 (d,1H, H indolizine); 7.6 (s, 1H, H indolizine); 7.15 (m, 2H, Hindolizine); 6.85 (m, 1H, H indolizine); 4.25 (q, 2H, —C(O)CH₂CH₃); 1.35(t, 3H, —C(O)CH₂CH₃)

IR: ν: C═O ester: 1675 cm⁻¹; aromatic C═C moieties: 1634 cm⁻¹

Step C: 4-Chloro-2-[1-(methoxycarbonyl)-3-indolizinyl]benzoic acid

The procedure is in accordance with the protocol described in Steps Cand D of Preparation 1, replacing the6-bromo-1,3-benzodioxole-5-carbaldehyde used in Step C with2-bromo-4-chlorobenzaldehyde.

Preparation 6:2-[2-(tert-Butoxycarbonyl)-8-(methoxycarbonyl)-1,2,3,4-tetrahydropyrrolo[1,2-a]pyrazin-6-yl]-4-fluorobenzoicacid

The procedure is in accordance with the protocol described inPreparation 2, replacing the 2-bromo-4-chlorobenzaldehyde used in Step Dwith 2-bromo-4-fluorobenzaldehyde.

Preparation 7:6-[2-(tert-Butoxycarbonyl)-8-(methoxycarbonyl)-1,2,3,4-tetrahydropyrrolo[1,2-a]pyrazin-6-yl]-1,3-benzodioxole-5-carboxylicacid

The procedure is in accordance with the protocol described inPreparation 2, replacing the 2-bromo-4-chlorobenzaldehyde used in Step Dwith 6-bromo-1,3-benzodioxole-5-carbaldehyde.

Preparation 8:6-[1′-(Methoxycarbonyl)-5′,6′-dihydro-8′H-spiro[1,3-dioxolane-2,7′-indolizin]-3′-yl]-1,3-benzodioxole-5-carboxylicacid Step A: Methyl8-formyl-1,4-dioxa-8-azaspiro[4.5]decane-7-carboxylate

24 g of methyl 1,4-dioxa-8-azaspiro[4.5]decane-9-carboxylate (111 mmol)are dissolved in 80 mL of ethyl acetate and 80 mL of dichloromethane.There are added 26 g of (4-nitrophenyl)formate (155 mmol) and the batchis stirred at ambient temperature for 1 hour. The reaction mixture isevaporated to dryness and taken up in ethyl acetate. The organic phaseis then successively washed with 1N NaOH solution, water and then withsaturated NH₄Cl solution until a neutral pH is obtained. It is thendried over magnesium sulphate, filtered and concentrated to dryness. Theoil thereby obtained is purified by flash chromatography (heptane/ethylacetate gradient). The title product is obtained in the form of an oil.

¹H NMR: δ (400 MHz; dmso-d6; 300K): 8.15 (s, 1H, CHO); 5.0-4.75 (m, 1H,tertiary H); 4.3-3.7 (m, 5H, 4H ethylenedioxy+1H aliphatic piperidine);3.70 (s, 3H, Me); 3.4-2.9 (2m, 1H, H aliphatic piperidine); 2.3-1.75 (m,2H, H aliphatic piperidine); 1.7-1.5 (m, 2H, H aliphatic piperidine)

Step B: 8-Formyl-1,4-dioxa-8-azaspiro[4.5]decane-7-carboxylic acid

15.25 g of the compound obtained in Step A (62.7 mmol) is dissolved in160 mL of dioxane. A solution of 125 mL of 1M KOH is added dropwise andthe batch is stirred at ambient temperature for 1 hour. There are thenadded 125 mL of 1M HCl and the compound is extracted withdichloromethane. The organic phase is dried over MgSO₄, filtered andconcentrated to dryness. The title product is obtained in the form of apowder.

¹H NMR: δ (400 MHz; dmso-d6; 300K) 13.5-12 (m, 1H, OH); 8.1+8.0 (2s, 1H,CHO); 4.9+4.6 (2m, 1H, tertiary H); 4.0-3.8 (m, 4H, ethylenedioxy);4.2+3.7 (2 ms, 1H, H aliphatic piperidine); 3.4+2.9 (2m, 1H, H aliphaticpiperidine); 2.4-1.5 (m, 4H, H aliphatic piperidine)

IR: ν: OH: 3500-2000 cm⁻¹; —C═O (acid+aldehyde): 1731+1655 cm⁻¹

Step C: Methyl 5′,6′-dihydro-8′H-spiro[1,3-dioxolane-2,7′-indolizine]-1′-carboxylate

To a solution of 13.5 g (62.7 mmol) of the acid obtained in Step B in380 mL of dichloromethane there are successively added 39.5 mL (238.4mmol) of triethylamine and then, in portions, 12.5 g (65.6 mmol) ofpara-toluenesulphonyl chloride and 23.7 mL (238.4 mmol) of methylchloroacrylate. The batch is stirred at 80° C. for 18 hours. Thereaction mixture is then filtered over Celite. The filtrate is thenwashed with saturated NaHCO₃ solution and then with saturated NH₄Clsolution. The organic phase is dried over MgSO₄, filtered andconcentrated to dryness. The oil thereby obtained is purified by flashchromatography (heptane/ethyl acetate gradient). The product is obtainedin the form of a solid.

¹H NMR: δ (400 MHz; dmso-d6; 300K) 6.70 (d, 1H, pyrrole); 6.40 (d, 1H,pyrrole); 4.05 (t, 2H, H aliphatic, piperidine); 4.00 (m, 4H,ethylenedioxy); 3.70 (s, 3H, methyl); 3.15 (s, 2H, H aliphaticpiperidine); 2.05 (t, 2H, H aliphatic piperidine)

IR: ν: —C═O (ester): 1689 cm⁻¹

Step D: Methyl3′-(6-formyl-1,3-benzodioxol-5-yl)-5′,6′-dihydro-8′H-spiro[1,3-dioxolane-2,7′-indolizine]-1′-carboxylate

The procedure is in accordance with the process of Step C of Preparation1.

Step E:6-[1′-(Methoxycarbonyl)-5′,6′-dihydro-8′H-spiro[1,3-dioxolane-2,7′-indolizin]-3′-yl]-1,3-benzodioxole-5-carboxylicacid

The procedure is in accordance with the process of Step D of Preparation1.

Preparation 9:6-[1-(Methoxycarbonyl)-3-indolizinyl]-1,3-benzodioxole-5-carboxylic acid

The procedure is in accordance with the protocol described inPreparation 5, replacing the 2-bromo-4-chlorobenzaldehyde used in Step Cwith 6-bromo-1,3-benzodioxole-5-carbaldehyde.

Preparation 10:4-Methyl-2-[1-(methoxycarbonyl)-5,6,7,8-tetrahydro-3-indolizinyl]-benzoicacid

The procedure is in accordance with the protocol described inPreparation 1, replacing the 6-bromo-1,3-benzodioxole-5-carbaldehydeused in Step C with 2-bromo-4-methylbenz-aldehyde.

Preparation 11:2-[1-(Methoxycarbonyl)-5,6,7,8-tetrahydro-3-indolizinyl]benzoic acid

The procedure is in accordance with the protocol described inPreparation 1, replacing the 6-bromo-1,3-benzodioxole-5-carbaldehydeused in Step C with 2-bromo-benzaldehyde.

Preparation 12:6-[8-(Methoxycarbonyl)pyrrolo[1,2-a]pyrimidin-6-yl]-1,3-benzodioxole-5-carboxylicacid Step A: Methyl pyrrolo[1,2-a]pyrimidine-8-carboxylate

To a solution of 6.2 g of methyl 2-pyrimidin-2-ylacetate (40.75 mmol) in250 mL of acetone there are successively added 14.04 g (167 mmol) ofNaHCO₃ in the form of a powder, 13.2 mL (203.75 mmol) ofchloroacetaldehyde and then 3.54 g (40.75 mmol) of lithium bromide. Thebatch is heated at 60° C. for 24 hours. The reaction mixture is thenconcentrated to dryness, taken up in ethyl acetate, washed with water,dried over MgSO₄, filtered and then concentrated to dryness. The solidthereby obtained is then purified by chromatography over silica gel(AcOEt). The expected product is obtained in the form of an oil.

Mass spectrum:

Empirical formula: C₈H₈N₂O₂

LC/MS: m/z=[M+H]⁺=177

Step B: Methyl6-(6-formyl-1,3-benzodioxol-5-yl)pyrrolo[1,2-a]pyrimidine-8-carboxylate

To a solution of 3.93 g of the compound obtained in Step A (22.3 mmol)in 80 mL of anhydrous dimethylacetamide there are added 7.66 g (33.45mmol) of 6-bromo-1,3-benzodioxole-5-carbaldehyde and 4.4 g (44.6 mmol)of potassium acetate. The batch is degassed under nitrogen for 15minutes. There are then added 1.56 g (2.23 mmol) of PdCl₂(PPh₃)₄catalyst. The reaction mixture is heated at 130° C. for 16 hours underan inert atmosphere. After drying, the residue is taken up indichloromethane; the batch is filtered over a cake of Celite and thenthe filtrate is washed with water, dried over MgSO₄ and concentrated todryness. The black solid is then chromatographed over silica gel(CH₂Cl₂/MeOH 5%). The expected product is obtained in the form of asolid.

Mass Spectrum:

Empirical formula: C₁₇H₁₂N₂O₃

LC/MS: m/z=[M+H]⁺=325

Step C:6-[8-(Methoxycarbonyl)pyrrolo[1,2-a]pyrimidin-6-yl]-1,3-benzodioxole-5-carboxylicacid

To a solution of 2.91 g (8.97 mmol) of the aldehyde obtained in Step Bin 140 mL of acetone cooled to 0° C. there are added 2-methylbutene andthen, dropwise, a mixture of 2.8 g (17.94 mmol) of NaH₂PO₄.2H₂O and 2.84g (31.4 mmol) of NaClO₂ dissolved in 30 mL of water. The batch isstirred at ambient temperature for 4 hours. The reaction mixture is thenconcentrated in vacuo to remove the acetone, placed at 0° C. and thenacidified to pH=2-3 by adding 5N HCl solution dropwise. The formation ofa precipitate is observed, which is filtered off, washed with water andthen with diethyl ether and dried in vacuo.

¹H NMR: δ (400 MHz; dmso-d6; 300K): 12.7 (m, 1H, COOH); 8.45 (d, 1H,aromatic H, H pyrrolo[1,2-a]pyrimidine); 8.19 (d, 1H, aromatic H, Hpyrrolo[1,2-a]pyrimidine); 6.9 (dd, 1H, aromatic H, Hpyrrolo[1,2-a]pyrimidine); 7.51 (s, 1H, aromatic H); 7.21 (s, 1H,aromatic H); 7.07 (s, 1H, aromatic H); 6.2 (s, 2H, aliphatic Hs,O—CH₂—O); 3.8 (s, 3H, aliphatic Hs, COOCH₃)

IR: ν —OH—: 3300 to 1800 cm⁻¹; ν —CO—: 1705 cm⁻¹, ν >C═C<: 1616 cm⁻¹

Preparation 13:4-Methoxy-2-[1-(methoxycarbonyl)-5,6,7,8-tetrahydro-3-indolizinyl]-benzoicacid

The procedure is in accordance with the protocol described inPreparation 1, replacing the 6-bromo-1,3-benzodioxole-5-carbaldehydeused in Step C with 2-bromo-4-methoxy-benzaldehyde.

Preparation 14:5-Methoxy-2-[1-(methoxycarbonyl)-5,6,7,8-tetrahydro-3-indolizinyl]-benzoicacid

The procedure is in accordance with the protocol described inPreparation 1, replacing the 6-bromo-1,3-benzodioxole-5-carbaldehydeused in Step C with 2-bromo-5-methoxy-benzaldehyde.

Preparation 15:7-[1-(Methoxycarbonyl)-5,6,7,8-tetrahydro-3-indolizinyl]-2,3-dihydro-1,4-benzodioxine-6-carboxylicacid

The procedure is in accordance with the process of Preparation 1,replacing the 6-bromo-1,3-benzodioxole-5-carbaldehyde used in Step Cwith 7-bromo-2,3-dihydro-1,4-benzodioxine-6-carbaldehyde.

Preparation 16: 2-[1-(Methoxycarbonyl)-3-indolizinyl]benzoic acid

The procedure is in accordance with the process of Preparation 5,replacing the 2-bromo-4-chlorobenzaldehyde used in Step C with2-bromo-benzaldehyde.

Preparation 17: 4-Fluoro-2-[1-(methoxycarbonyl)-3-indolizinyl]benzoicacid

The procedure is in accordance with the process of Preparation 5,replacing the 2-bromo-4-chlorobenzaldehyde used in Step C with2-bromo-4-fluorobenzaldehyde.

Preparation 18:4-Fluoro-2-[1-(methoxycarbonyl)-5,6,7,8-tetrahydro-3-indolizinyl]-benzoicacid

The procedure is in accordance with the process of Preparation 1,replacing the 6-bromo-1,3-benzodioxole-5-carbaldehyde used in Step Cwith 2-bromo-4-fluorobenzaldehyde.

Preparation 1′: (3R)-3-Methyl-1,2,3,4-tetrahydroisoquinolinehydrochloride Step A:{(3S)-2-[(4-Methylphenyl)sulphonyl]-1,2,3,4-tetrahydroisoquinolin-3-yl}methyl4-methylbenzenesulphonate

To a solution of 30.2 g of[(3S)-1,2,3,4-tetrahydroisoquinolin-3-yl]methanol (185 mmol) in 750 mLof dichloromethane there are successively added 91.71 g of tosylchloride (481 mmol) and then, dropwise, 122.3 mL of N,N,N-triethylamine(740 mmol). The reaction mixture is then stirred at ambient temperaturefor 20 hours. It is then diluted with dichloromethane, washedsuccessively with 1M HCl solution, saturated NaHCO₃ solution and thensaturated NaCl solution until neutral. The organic phase is then driedover MgSO₄, filtered and concentrated to dryness. The solid obtained isthen dissolved in a minimum volume of dichloromethane and thencyclohexane is added until a precipitate is formed. This precipitate isthen filtered off and washed with cyclohexane. After drying, the titleproduct is obtained in the form of crystals.

¹H NMR: δ (400 MHz; dmso-d6; 300K): 7.75 (d, 2H, aromatic Hs, orthoO-tosyl); 7.6 (d, 2H, aromatic Hs, ortho N-tosyl); 7.5 (d, 2H, aromaticHs, meta O-tosyl); 7.3 (d, 2H, aromatic Hs, meta N-tosyl); 7.15-6.9 (m,4H, aromatic Hs, tetrahydroisoquinoline); 4.4-4.15 (dd, 2H, aliphaticHs, tetrahydroisoquinoline); 4.25 (m, 1H, aliphatic H,tetrahydroisoquinoline); 4.0-3.8 (2dd, 2H, aliphatic Hs, CH₂—O-tosyl);2.7 (2dd, 2H, aliphatic Hs, tetrahydroisoquinoline); 2.45 (s, 3H,O—SO₂-Ph-CH₃); 2.35 (s, 3H, N—SO₂-Ph-CH₃)

IR: ν: —SO₂: 1339-1165 cm⁻¹

Step B:(3R)-3-Methyl-2-[(4-methylphenyl)sulphonyl]-1,2,3,4-tetrahydroisoquinoline

To a suspension of 8.15 g (214.8 mmol) of LiAlH₄ in 800 mL of methyltert-butyl ether (MTBE) there are added 101.2 g of the ditosyl compoundobtained in Step A (214.8 mmol) dissolved in 200 mL of MTBE. The batchis then heated at 50° C. for 2 hours. It is allowed to cool and placedat 0° C., and there are then added, dropwise, 12 mL of 5N NaOH solution.The batch is stirred at ambient temperature for 45 minutes. The solidthereby obtained is then filtered off and washed with MTBE and then withdichloromethane. The filtrate is then concentrated to dryness. The titleproduct is then obtained in the form of a solid.

¹H NMR: δ (400 MHz; dmso-d6; 300K): 7.70 (d, 2H, aromatic Hs, orthoN-tosyl); 7.38 (d, 2H, aromatic Hs, meta N-tosyl); 7.2-7.0 (m, 4H,aromatic Hs, tetrahydroisoquinoline); 4.4 (m, 2H, aliphatic Hs,tetrahydroisoquinoline); 4.3 (m, 1H, aliphatic H,tetrahydroisoquinoline); 2.85-2.51 (2dd, 2H, aliphatic Hs,tetrahydroisoquinoline); 2.35 (s, 3H, N—SO₂-Ph-CH₃); 0.90 (d, 3H,tetrahydroisoquinoline-CH₃)

IR: ν: —SO₂: 1332-1154 cm⁻¹

Step C: (3R)-3-Methyl-1,2,3,4-tetrahydroisoquinoline

To a solution of 31.15 g (103.15 mmol) of the monotosyl compoundobtained in Step B in 500 mL of anhydrous methanol there are added, inportions, 3.92 g (161 mmol) of magnesium turnings. The batch is stirredin the presence of ultrasound for 96 hours. The reaction mixture is thenfiltered and the solid is washed several times with methanol. Thefiltrate is then concentrated to dryness. After purification by columnchromatography over silica gel (dichloromethane/EtOH/NH₄OH), the titleproduct is obtained in the form of an oil.

¹H NMR: δ (400 MHz; dmso-d6; 300K): 7.05 (m, 4H, aromatic Hs,tetrahydroisoquinoline); 3.90 (m, 2H, aliphatic Hs,tetrahydroisoquinoline); 2.85 (m, 1H, aliphatic H,tetrahydroisoquinoline); 2.68-2.4 (2dd, 2H, aliphatic Hs,tetrahydroisoquinoline); 1.12 (d, 3H, tetrahydroisoquinoline-CH₃);2.9-2.3 (m, broad, 1H, HN (tetrahydroisoquinoline))

IR: ν: —NH: 3248 cm⁻¹

Step D: (3R)-3-Methyl-1,2,3,4-tetrahydroisoquinoline hydrochloride

To a solution of 14.3 g (97.20 mmol) of the compound obtained in Step Cin 20 mL of anhydrous ethanol there are added, dropwise, 100 mL of a 1Msolution of HCl in ether. The batch is stirred at ambient temperaturefor 1 hour and then filtered. The crystals thereby obtained are washedwith ethyl ether. After drying, the title product is obtained in theform of crystals.

¹H NMR: δ (400 MHz; dmso-d6; 300K): 9.57 (m, broad, 2H, NH₂ ⁺(tetrahydroisoquinoline); 7.22 (m, 4H, aromatic Hs,tetrahydroisoquinoline); 4.27 (s, 2H, aliphatic Hs,tetrahydroisoquinoline); 3.52 (m, 1H, aliphatic H,tetrahydroisoquinoline); 3.03-2.85 (2dd, 2H, aliphatic Hs,tetrahydroisoquinoline); 1.39 (d, 3H, tetrahydroisoquinoline-CH₃)

IR: ν: —NH₂+: 3000-2300 cm⁻¹; ν: aromatic —CH: 766 cm⁻¹

Preparation 2′:(3S)-3-[2-(Morpholin-4-yl)ethyl]-1,2,3,4-tetrahydroisoquinolinehydrochloride Step A: tert-Butyl(3S)-3-(2-morpholino-2-oxo-ethyl)-3,4-dihydro-1H-isoquinoline-2-carboxylate

To a solution of 3 g (10.30 mmol) of[(3S)-2-(tert-butoxycarbonyl)-1,2,3,4-tetrahydroisoquinolin-3-yl]aceticacid in 100 mL of dichloromethane there are added, dropwise, 1.10 mL(11.32 mmol) of morpholine and, still dropwise, 4.3 mL (30.9 mmol) oftriethylamine, 2.20 g (12.40 mmol) of 1,2-dichloromethane and 1.70 g(1.68 mmol) of hydroxybenzotriazole. The batch is stirred at ambienttemperature for 15 hours. The reaction mixture is then diluted withdichloromethane and washed successively with 1M HCl solution, saturatedNaHCO₃ solution and then saturated NaCl solution until neutral. Theorganic phase is then dried over MgSO₄, filtered and concentrated todryness. After purification by column chromatography over silica gel(dichloromethane/MeOH), the title product is obtained in the form of anoil.

¹H NMR: δ (400 MHz; dmso-d6; 300K): 7.20-7.10 (m, 4H, aromatic Hs,tetrahydroisoquinoline); 4.70 (m, 1H, aliphatic Hs, CHtetrahydroisoquinoline); 4.75-4.20 (2m, 2H, aliphatic Hs, CH₂ alpha to Ntetrahydroisoquinoline); 3.60 (m, 8H, aliphatic Hs, morpholine); 3.00and 2.70 (2dd, 2H, aliphatic H, tetrahydroisoquinoline); 2.50-2.20 (2d,2H, aliphatic Hs, CH₂CO); 1.40 (s, 9H, ^(t)Bu)

IR: ν: C═O: 1687; 1625 cm⁻¹

Step B:1-(Morpholin-4-yl)-2-[(3S)-1,2,3,4-tetrahydroisoquinolin-3-yl]ethanonehydrochloride

To a solution of 2.88 g (7.18 mmol) of the compound obtained in Step Ain 16 mL of dichloromethane there are added, dropwise, 80 mL (80 mmol)of a 1M solution of HCl in ether. The batch is stirred at ambienttemperature for 15 hours and then the suspension is filtered and theprecipitate is washed with ether. After drying, the title product isobtained in the form of a solid.

¹H NMR: δ (400 MHz; dmso-d6; 300K): 9.80-9.50 (m, 2H, NH₂ ⁺); 7.30-7.10(m, 4H, aromatic Hs, tetrahydroisoquinoline); 4.30 (m, 2H, aliphatic Hs,CH₂ alpha to N tetrahydroisoquinoline); 3.80 (m, 1H, aliphatic Hs, CHtetrahydroisoquinoline); 3.70-3.40 (2m, 8H, aliphatic Hs, morpholine);3.15 and 2.8 (m, 4H, aliphatic H, CH₂ tetrahydroisoquinoline and CH₂CO)

IR: ν: —NH₂+: 2800-1900 cm⁻¹; ν: C═O: 1620 cm⁻¹

Step C: (3S)-3-[2-(Morpholin-4-yl)ethyl]-1,2,3,4-tetrahydroisoquinolinehydrochloride

A solution of 2.2 g (7.44 mmol) of the compound obtained in Step B in 22mL of MTBE and 5 mL of dichloromethane is prepared. After cooling in anice bath at 0° C., there are added thereto, dropwise, 15 mL (15 mmol) of1M LiAlH₄ solution in tetrahydrofuran. The batch is then stirred atambient temperature for 6 hours. It is placed at 0° C., and there isthen added, dropwise, 1 mL of 5N NaOH solution. The batch is stirred atambient temperature for 45 minutes. The solid is then filtered off andwashed with MTBE and then with dichloromethane and the filtrate isconcentrated to dryness. The oil thereby obtained is diluted withdichloromethane and there are added, dropwise, 6.3 mL of a 1M solutionof HCl in ether. The batch is stirred at ambient temperature for 1 hourand then filtered. The crystals thereby obtained are washed with ethylether. After drying, the title product is obtained in the form of asolid.

¹H NMR: δ (400 MHz; dmso-d6; 300K): 11.35+9.80 (2m, 2H, NH₂ ⁺); 10.00(m, H, NH⁺); 7.20 (m, 4H, aromatic Hs, tetrahydroisoquinoline); 4.30 (s,2H, aliphatic Hs, CH₂ alpha to N tetrahydroisoquinoline); 4.00+3.85 (2m,4H, aliphatic Hs, CH₂ alpha to N morpholine); 3.70 (m, 1H, aliphatic Hs,CH tetrahydroisoquinoline); 3.55-3.30 (m, 4H, aliphatic Hs, CH alpha toO morpholine and CH₂-morpholine); 3.15 (dd, 1H, aliphatic H, CH₂tetrahydroisoquinoline); 3.10 (m, 2H, aliphatic H, CH alpha to 0morpholine); 2.90 (dd, 1H, aliphatic H, CH₂ tetrahydroisoquinoline);2.30+2.15 (2m, 2H, aliphatic H, CH₂-tetrahydroisoquinoline)

IR: ν: NH⁺/—NH₂+: between 3500 and 2250 cm⁻¹; ν: C═C: weak 1593 cm⁻¹; ν:aromatic C—H: 765 cm⁻¹

Preparation 3′: tert-Butyl{2-[(3S)-1,2,3,4-tetrahydroisoquinolin-3-yl]ethyl}-carbamate Step A:Benzyl (3S)-3-(2-hydroxyethyl)-3,4-dihydroisoquinoline-2(1H)-carboxylate

The title compound is obtained starting from(3S)-2-[(benzyloxy)carbonyl]-1,2,3,4-tetrahydroisoquinoline-3-carboxylicacid based on a protocol from the literature (Jinlong Jiang et alBioorganic & Medicinal Chemistry Letters, 14, 1795, 2004).

Step B: Benzyl(3S)-3-{2-[(methylsulphonyl)oxy]ethyl}-3,4-dihydroisoquinoline-2(1H)-carboxylate

To a solution of 10.6 g of the compound of Step A (35.6 mmol) in 350 mLof anhydrous CH₂Cl₂ placed at 0° C. there are successively added 10.1 mLof triethylamine (71.2 mmol) and then, dropwise, 3.1 mL ofmethanesulphonyl chloride (39 mmol). The reaction mixture is thenstirred at ambient temperature for 2 hours. Hydrolysis is then carriedout by slowly adding water. The product is extracted several times withCH₂Cl₂. The organic phases are then combined and successively washedwith 1N HCl solution, saturated NaCl solution, saturated NaHCO₃ solutionand saturated NaCl solution until neutral. They are then dried overMgSO₄ and concentrated to dryness. After purification by chromatographyover silica gel (petroleum ether/AcOEt gradient), the expected productis obtained in the form of a foam.

LC/MS: m/z=(M+H)⁺=375

Step C: Benzyl(3S)-3-(cyanomethyl)-3,4-dihydroisoquinoline-2(1H)-carboxylate

To a solution of 15.4 g of the compound obtained in Step B (41.02 mmol)in 250 mL of anhydrous DMSO there are added 22 g (449 mmol) of sodiumcyanide. The batch is then heated at 60° C. for 12 hours. It is allowedto cool and then the reaction mixture is diluted by adding ethylacetate. Hydrolysis is then carried out with saturated NaHCO₃ solution.After extracting two more times with ethyl acetate, the organic phasesare combined, washed with H₂O, dried over MgSO₄ and concentrated todryness. After purification by chromatography over silica gel(hexane/AcOEt 7/3), the expected product is obtained in the form of anoil.

LC/MS: m/z=[M+H]⁺=307.1

Step D: Benzyl(3S)-3-(2-aminoethyl)-3,4-dihydroisoquinoline-2(1H)-carboxylate

To a solution of 15.4 g of the compound obtained in Step C (50.3 mmol)in 300 mL of anhydrous THF placed at 0° C. there are added, dropwise, a1N solution of BH₃-THF. The reaction mixture is allowed to come back toambient temperature gradually and then the batch is stirred for 14hours. The reaction mixture is then hydrolysed by slowly addingsaturated NH₄Cl solution. After extracting twice with ethyl acetate, theorganic phases are combined and dried over MgSO₄. After concentrating todryness, the expected product is obtained in the form of a foam which isused directly without purification in the next protection step.

Step E: Benzyl(3S)-3-{2-[(tert-butoxycarbonyl)amino]ethyl}-3,4-dihydroisoquinoline-2(1H)-carboxylate

To a solution of 15.6 g of the compound obtained in Step D (50.3 mmol)in 670 mL of CH₂Cl₂ there are successively added, in portions, 13.2 g(60.36 mmol) of Boc₂O, 14 mL of triethylamine (100.6 mmol) and DMAP in acatalytic amount. The batch is stirred at ambient temperature for 5hours. The reaction mixture is then hydrolysed with water and extractedtwice with CH₂Cl₂. The organic phases are combined, washed with waterand dried over MgSO₄. After concentration to dryness and purification bychromatography over silica gel (heptane/AcOEt gradient), the expectedproduct is obtained in the form of an oil.

LC/MS: m/z=(M+H)⁺=411

Step F: tert-Butyl{2-[(3S)-1,2,3,4-tetrahydroisoquinolin-3-yl]ethyl}carbamate

To a solution of 10.4 g of the compound obtained in Step E (25.5 mmol)in 210 mL of anhydrous MeOH there are added 2.71 g (2.55 mmol) of Pd/C10%. The batch is degassed for 30 minutes and is then stirred under ahydrogen atmosphere for 16 hours. The reaction mixture is then filteredand concentrated to dryness. The expected product is obtained in theform of a solid which is taken up in a mixture of pentane/Et₂O (90/10),triturated and filtered. After drying, the product is obtained in theform of a solid.

¹H NMR: δ (400 MHz; dmso-d6; 300K): 7.1-6.98 (m, 4H, aromatic Hs,tetrahydroisoquinoline); 6.83 (m, 1H, CH₂NHBoc); 3.85 (s, 2H, aliphaticHs, tetrahydroisoquinoline); 3.09 (q, 2H, CH₂NHBoc); 2.73 (m, 1H,aliphatic Hs, tetrahydroisoquinoline); 2.70 and 2.39 (2m, 2H, aliphaticHs, tetrahydroisoquinoline); 1.63 (m, 2H, aliphatic Hs); 1.38 (s, 9H,NHCOOtBu)

IR: ν: >NH: 3378, −3201 cm⁻¹ (amine, amide); ν: >C═O: 1683 cm⁻¹ (amide);ν: >NH: 1524 cm⁻¹ (amide); ν: >C═O: 1168 cm⁻¹

LC/MS: m/z=[M+H]⁺=277

Preparation 4′:(3R)-3-[3-(Morpholin-4-yl)propyl]-1,2,3,4-tetrahydroisoquinoline Step A:{(3S)-2-[(4-Methylphenyl)sulphonyl]-1,2,3,4-tetrahydroisoquinolin-3-yl}methyl4-methylbenzenesulphonate

The procedure is the same as that of Step A of Preparation 1′.

Step B: tert-Butyl2-({(3R)-2-[(4-methylphenyl)sulphonyl]-1,2,3,4-tetrahydroisoquinolin-3-yl}methyl)-3-(morpholin-4-yl)-3-oxopropanoate

To a suspension of 1 g of NaH (60%) (25.08 mmol) in 30 mL of MTBE thereare added, dropwise, a solution of 5 g of tert-butyl3-morpholino-3-oxopropanoate (21.81 mmol) in 20 mL of anhydrous MTBE.This suspension stirred at ambient temperature for 1 hour and then thecompound obtained in Step A is added in the form of a powder. The batchis stirred at 60° C. for 30 hours. 100 mL of saturated ammonium chloridesolution are added. The resulting solution is extracted withdichloromethane. The organic phase is then dried over MgSO₄, filteredand concentrated to dryness. After purification by column chromatographyover silica gel (dichloromethane/MeOH), the expected product is obtainedin the form of an oil.

¹H NMR (500 MHz; dmso-d6) δ ppm: 7.63/7.59 (2d, 2H), 7.3/7.26 (2d, 2H);7.13 (m, 2H), 7.09/6.97 (2t, 2H), 4.64/4.55/4.36/4.28 (2AB, 2H),4.25/4.11 (2m, 1H), 3.81 (m, 1H), 3.73/3.48 (m, 4H), 3.57-3.32 (m, 4H),2.51 (m, 2H), 2.32/2.31 (2s, 3H), 1.88/1.79 (2m, 2H), 1.39/1.38 (2s,9H).

IR (ATR) cm⁻¹: ν: >C═O: 1731 (ester); ν: >C═O: 1644 (amide); ν: —SO2:1334-1156; ν: >C—O—C<: 1155; γ: >CH—Ar: 815-746-709

Step C:2-({(3R)-2-[(4-Methylphenyl)sulphonyl]-1,2,3,4-tetrahydroisoquinolin-3-yl}methyl)-3-(morpholin-4-yl)-3-oxopropanoicacid

To a solution of 9.5 g (17.97 mmol) of the compound obtained in Step Bin 40 mL of dioxane there are added, dropwise, 20 mL of a 4M solution ofHCl in dioxane. The batch is stirred at ambient temperature for 48 hoursand then the solution is concentrated to dryness. After drying, theexpected product is obtained in the form of an oil.

¹H NMR (400 MHz; dmso-d6) δ ppm: 12.75 (m, 1H), 7.6 (2*d, 2H); 7.3 (2*d,2H), 7.1/6.95 (2*m, 4H), 4.7/4.2 (d, 2H), 4.25/4.12 (2*m, 1H), 3.9-3.3(m, 9H), 2.55 (d, 2H), 2.3 (2*s, 3H), 1.8 (t, 2H)

IR (ATR) cm⁻¹: ν: —OH: 3500 to 2000; ν: >C═O: 1727 (acid); ν: >C═O: 1634(amide); ν: —SO2: 1330-1155

Step D:3-{(3R)-2-[(4-Methylphenyl)sulphonyl]-1,2,3,4-tetrahydroisoquinolin-3-yl}-1-(morpholin-4-yl)propan-1-one

To a solution of 7.80 g (16.51 mmol) of the compound obtained in Step Cin 100 mL of DMSO there are added 1.16 g (19.83 mmol) of solid sodiumchloride and then, dropwise, 5 mL of water. The batch is stirred at 130°C. for 1 hour and then the solution is concentrated to 3/4. The reactionmixture is then diluted with dichloromethane and washed successivelywith saturated lithium chloride solution and then with saturated NaClsolution. The organic phase is then dried over MgSO₄, filtered andconcentrated to dryness. After purification by column chromatographyover silica gel (cyclohexane/ethyl acetate), the expected product isobtained in the form of an oil.

¹H NMR (400 MHz; dmso-d6) δ ppm: 7.65 (d, 2H), 7.3 (d, 2H); 7.15/7 (2m,4H), 4.6 (d, 1H), 4.25 (d, 1H), 4.2 (m, 1H), 3.5 (m, 4H), 3.4 (2 m, 4H),2.6 (2 dd, 2H), 2.35 (s, 3H), 2.3 (m, 2H), 1.5 (quad., 2H)

IR (ATR) cm⁻¹: ν: >C═O: 1639; ν: —SO2: 1331-1156; γ: >CH—Ar: 815-675

Step E:(3R)-2-[(4-Methylphenyl)sulphonyl]-3-[3-(morpholin-4-yl)propyl]-1,2,3,4-tetrahydroisoquinoline

To a solution of 6.0 g (14.0 mmol) of the compound obtained in Step D in60 mL of MTBE and 14 mL of dichloromethane there are added 1.06 g (28mmol) of LAH in portions over 5 minutes. The batch is stirred at ambienttemperature for 15 hours. There are added, dropwise, 1.5 mL of water andstirring is carried out for 15 minutes. There are then added, dropwise,1.5 mL of 5M sodium hydroxide solution and stirring is carried out for15 minutes. The reaction mixture is then diluted with MTBE anddichloromethane. The suspension is then filtered and the precipitate iswashed with MTBE and dichloromethane. The organic phase is then driedover MgSO₄, filtered and concentrated to dryness. After purification bycolumn chromatography over silica gel (dichloromethane/EtOH/NH₄OH), theexpected product is obtained in the form of an oil.

¹H NMR (400 MHz; dmso-d6) δ ppm: 7.68 (d, 2H), 7.32 (d, 2H); 7.1(unresolved peak, 4H), 4.65/4.23 (AB, 2H), 4.2 (m, 1H), 3.55 (t, 4H),2.7/2.6 (ABx, 2H), 2.35 (s, 3H), 2.25 (t, 4H), 2.2 (t, 2H), 1.4/1.3 (2m,4H)

IR (ATR) cm⁻¹: ν: —SO2: 1333-1158

Step F: (3R)-3-[3-(Morpholin-4-yl)propyl]-1,2,3,4-tetrahydroisoquinoline

To a solution of 1.50 g (3.62 mmol) of the compound obtained in Step Ein 20 mL of anhydrous methanol there are added 2.0 g (82.3 mmol), inportions, of magnesium turnings. The batch is stirred in the presence ofultrasound for 96 hours. The reaction mixture is then filtered, thesolid is washed several times with methanol, and the filtrate isconcentrated to dryness. After purification by column chromatographyover silica gel (dichloromethane/EtOH/NH₄OH), the expected product isobtained in the form of an oil.

¹H NMR (400 MHz; dmso-d6) δ ppm: 7.3 (d, 2H), 7.1 (t, 2H); 7.1 (d+t,3H), 7 (d, 2H), 3.9 (s, 2H), 3.55 (t, 4H), 2.75 (m, 1H), 2.72/2.45 (dd,2H), 2.35 (t, 4H), 2.25 (t, 2H), 1.6 (m, 2H), 1.45 (m, 2H)

IR (ATR) cm⁻¹: ν: >NH2+/NH+: 3500-2300; ν: >C—O—C<: 1115

High-Resolution Mass Spectroscopy (ESI+−/FIA/HR):

Empirical formula: C₁₆H₂₄N₂O

[M+H]⁺, calculated: 261.1961

[M+H]⁺, measured: 261.1959

Preparation 5′:(3S)-3-[2-(3,3-Difluoropiperidin-1-yl)ethyl]-1,2,3,4-tetrahydroisoquinolinehydrochloride

The procedure is in accordance with the process of Preparation 2′,replacing the morpholine used in Step A with 3,3-difluoro-1-piperidine.

¹H NMR: δ (400 MHz; dmso-d6; 300K): 11.3 (m, 1H, NH⁺); 10.2-9.8 (m, 2H,NH₂ ⁺); 7.25 (m, 4H, aromatic Hs, tetrahydroisoquinoline); 4.3 (broad s,2H, aliphatic Hs, CH tetrahydroisoquinoline); 4.0-3.3 (m, 7H, aliphaticHs); 3.15-2.95 (dd, 2H, aliphatic Hs, CH tetrahydroisoquinoline);2.4-1.9 (m, 6H, aliphatic Hs, H 3,3-difluoro-1-piperidine)

IR: ν: NH⁺/NH₂ ⁺: between 300 and 2500 cm⁻¹; ν: C—F: 1204 cm⁻¹

Preparation 6′:(3S)-3-[2-(3-Methoxyazetidin-1-yl)ethyl]-1,2,3,4-tetrahydroisoquinolinehydrochloride

The procedure is in accordance with the process of Preparation 2′,replacing the morpholine used in Step A with 3-methoxyazetidine.

¹H NMR: δ (400 MHz; dmso-d6; 300K): 11.3 (m, 1H, NH⁺); 10.00 (m, 2H, NH₂⁺); 7.20 (m, 4H, aromatic Hs, tetrahydroisoquinoline); 4.4 (m, 1H,aliphatic H, 3-methoxy azetidine); 4.30 (s, 2H, aliphatic Hs,tetrahydroisoquinoline); 4.2-3.45 (m, 4H, 3-methoxyazetidine); 4.2-3.6(m, 3H, aliphatic Hs); 3.1 and 2.95 (dd, 2H, aliphatic Hs); 3.25 (s, 3H,OCH₃)

Preparation 7′: (3S)-3-Methyl-1,2,3,4-tetrahydroisoquinolinehydrochloride

The procedure is in accordance with the process of Preparation 1′,replacing the [(3S)-1,2,3,4-tetrahydroisoquinolin-3-yl]methanol used inStep A with [(3R)-1,2,3,4-tetrahydroisoquinolin-3-yl]methanol.

Preparation 1″:N-(4-{[tert-Butyl(dimethyl)silyl]oxy}phenyl)-1-methyl-1H-pyrazol-4-amineStep A: 4-{[tert-Butyl(dimethyl)silyl]oxy}aniline

The title compound is obtained starting from 4-aminophenol in THF in thepresence of imidazole and tert-butyl(dimethyl)silyl chloride inaccordance with the protocol described in the literature (S. Knaggs etal, Organic & Biomolecular Chemistry, 3(21), 4002-4010; 2005).

¹H NMR: δ (400 MHz; dmso-d6; 300K): 6.45-6.55 (dd, 4H, aromatic Hs);4.60 (m, 2H, NH₂-Ph); 0.90 (s, 9H, Si (CH₂)₂CH(CH₃)₂); 0.10 (s, 6H, Si(CH₂)₂CH(CH₃)₂)

IR: ν: —NH₂+: 3300-3400 cm⁻¹

Step B:N-[4-[tert-Butyl(dimethyl)silyl]oxyphenyl]-1-methyl-pyrazol-4-amine

To a solution of 30.8 g (0.137 mol) of the compound of Step A in 525 mLof anhydrous toluene there are successively added 29.8 g of sodiumtert-butylate (0.310 mol), 4.55 g of Pd₂(dba)₃ (also referred to astris(dibenzylideneacetone)dipalladium(0)) (4.96 mmol), 4.81 g of2-di-tert-butylphosphino-2′,4′,6′-tri-isopropyl-1,1′-biphenyl (9.91mmol) and 12.8 mL of 4-bromo-1-methyl-1H-pyrazole (0.124 mol). The batchis degassed under argon for 30 minutes and then refluxed for 3 hours. Itis allowed to cool. The reaction mixture is concentrated to dryness andthen taken up in dichloromethane, filtered over Celite and thenconcentrated to dryness again. The residue is then purified bychromatography over silica gel (gradient CH₂Cl₂/AcOEt) to provide theexpected product in the form of a solid.

¹H NMR: δ (400 MHz; dmso-d6; 300K): 7.55 (s, 1H, pyrazole); 7.23 (s, 1H,pyrazole); 7.18 (broad s, 1H, NH₂-Ph); 6.64 (m, 4H, aromatic Hs); 3.77(s, 3H, CH₃-pyrazole); 0.90 (s, 9H, Si (CH₂)₂CH(CH₃)₂); 0.12 (s, 6H, Si(CH₂)₂CH(CH₃)₂)

IR: ν —NH⁺: 3275 cm⁻¹; ν Ar and C═N: 1577 and 1502 cm⁻¹; ν —Si—C—: 1236cm⁻¹; ν —Si—O—: 898 cm⁻¹; ν —Si—C—: 828, 774 cm⁻¹

Preparation 2″:N-(4-{[tert-Butyl(dimethyl)silyl]oxy}phenyl)-1-methyl-1H-indol-5-amine

The procedure is in accordance with the process of Preparation 1″,replacing the 4-bromo-1-methyl-1H-pyrazole used in Step B with5-bromo-1-methyl-1H-indole.

Preparation 3″:N-(4-{[tert-Butyl(dimethyl)silyl]oxy}phenyl)-1-[2-(morpholin-4-yl)ethyl]-1H-indol-5-amineStep A: 5-Bromo-1-[2-(morpholin-4-yl)ethyl]-1H-indole

To a suspension of NaH (4.5 g; 112 mmol) in anhydrous THF (300 mL)placed at 0° C. there are added, in portions, 5-bromo-1H-indole (10.4 g;51 mmol). After stirring for 20 minutes at 0° C.,4-(2-chloroethyl)morpholine hydrochloride (10.4 g; 56 mmol) is added inportions over 1 hour. After stirring overnight at ambient temperature,the reaction mixture is placed at 80° C. for 5 hours. It is then pouredover a mixture of aqueous sodium bicarbonate and dichloromethane. Theaqueous phase is extracted with dichloromethane. The organic phase isdried over MgSO₄ and concentrated to dryness, and the residue ispurified by chromatography over silica gel (CH₂Cl₂/MeOH gradient) toprovide the expected product in the form of an oil.

¹H NMR: δ (400 MHz; CDCl3; 300K): 7.75 (d, 1H); 7.30 (dd, 1H); 7.20 (d,1H); 7.15 (d, 1H); 6.40 (d, 1H); 4.20 (t, 2H); 3.70 (m, 4H); 2.75 (t,2H); 2.45 (m, 4H)

Step B: 5-Bromo-1-[2-(morpholin-4-yl)ethyl]-1H-indole

The procedure is in accordance with the process of Preparation 1″,replacing the 4-bromo-1-methyl-1H-pyrazole used in Step B with thecompound obtained in Step A.

¹H NMR: δ (400 MHz; dmso-d6; 300K): 7.35 (d, 1H); 7.15 (s, 1H); 6.85 (d,3H); 6.70 (d, 2H); 7.30 (d, 1H); 6.25 (d, 1H), 4.20 (t, 2H); 3.55 (m,4H); 2.65 (t, 2H); 2.45 (m, 4H); 1.45 (s, 9H), 0.15 (s, 6H)

Preparation 4″:N-(4-{[tert-Butyl(dimethyl)silyl]oxy}phenyl)-1-[2-(morpholin-4-yl)ethyl]-2,3-dihydro-1H-indol-5-amine

The procedure is in accordance with the process of Preparation 2″,replacing the 5-bromoindole used in Step A with5-bromo-2,3-dihydro-1H-indole.

Preparation 5″:N-(4-{[tert-Butyl(dimethyl)silyl]oxy}phenyl)-4-fluoroaniline

The procedure is in accordance with the process of Preparation 1″,replacing the 4-bromo-1-methyl-1H-pyrazole used in Step B with1-bromo-4-fluorobenzene.

Preparation 6″:N-(4-{[tert-Butyl(dimethyl)silyl]oxy}phenyl)-3-fluoro-4-methylaniline

The procedure is in accordance with the process of Preparation 1″,replacing the 4-bromo-1-methyl-1H-pyrazole used in Step B with4-bromo-2-fluoro-1-methylbenzene.

Preparation 7″:N-(4-{[tert-Butyl(dimethyl)silyl]oxy}phenyl)-1-methyl-1H-indazol-5-amine

The procedure is in accordance with the process of Preparation 1″,replacing the 4-bromo-1-methyl-1H-pyrazole used in Step B with5-bromo-1-methyl-1H-indazole.

Preparation 8″: 4-{[tert-Butyl(dimethyl)silyl]oxy}-N-phenylaniline

To a solution of 12 g of 4-anilinophenol (64.7 mmol) in 200 mL ofacetonitrile there are added, at ambient temperature, 6.7 g of imidazole(97.05 mmol) and 11.7 g of tert-butyl-(chloro)dimethylsilane (77.64mmol). The batch is stirred at 70° C. for 4 hours. The reaction mixtureis then poured into water and extracted with ether. The organic phase isthen dried over magnesium sulphate, then filtered and evaporated todryness. The crude product thereby obtained is then purified bychromatography over silica gel (petroleum ether/dichloromethanegradient). The title product is obtained in the form of a powder.

¹H NMR: δ (400 MHz; dmso-d6; 300K): 7.84 (s, 1H NH); 7.17 (t, 2Haniline); 6.98 (d, 2H phenoxy); 6.94 (d, 2H aniline); 6.76 (d, 2Hphenoxy); 6.72 (t, 1H aniline); 0.95 (s, 9H tert-butyl); 0.15 (s, 6Hdimethyl)

IR: ν: >NH: 3403 cm⁻¹; >Ar: 1597 cm⁻¹

Preparation 9″: 4-Benzyloxy-N-phenyl-aniline

To a solution of 4-hydroxy-N-phenyl-aniline (30 g; 162 mmol) inacetonitrile (400 mL) there are added 58 g of Cs₂CO₃ (178 mmol) andstirring is carried out for 15 minutes at ambient temperature. Benzylbromide (22.5 mL; 178 mmol) is then added dropwise and then the reactionmixture is refluxed for 4 hours. After filtering and rinsing withacetonitrile, the filtrate is concentrated and chromatographed oversilica gel (petroleum ether/AcOEt gradient). The title product is thenobtained in the form of a colourless solid.

Preparation 10″:N-(4-{[tert-Butyl(dimethyl)silyl]oxy}phenyl)-3-fluoro-4-[2-(morpholin-4-yl)ethoxy]aniline

The procedure is in accordance with the process of Preparation 3″,replacing the 5-bromo-1H-indole used in Step A by4-bromo-2-fluorophenol.

¹H NMR: δ (400 MHz; dmso-d6; 300K): 7.75 (d, 1H); 7 (dd, 1H); 6.9 (d,2H); 6.75 (m, 3H); 6.7 (ddd, 1H); 4.05 (t, 2H); 3.6 (t, 4H); 2.65 (t,2H); 2.45 (t, 4H); 0.95 (s, 9H); 0.2 (s, 6H)

Preparation 11″:N-(4-{[tert-Butyl(dimethyl)silyl]oxy}phenyl)pyridin-4-amine

The procedure is in accordance with the process of Preparation 1″,replacing the 4-bromo-1-methyl-1H-pyrazole used in Step B with4-bromopyridine.

IR: ν —NH—: 3200 and 2500 cm⁻¹; ν —Si—O—: 902 cm⁻¹; ν —Si—C—: 820 cm⁻¹

Preparation 12″:3-[(4-{[tert-Butyl(dimethyl)silyl]oxy}phenyl)amino]benzonitrile

The procedure is in accordance with the process of Preparation 1″,replacing the 4-bromo-1-methyl-1H-pyrazole used in Step B with3-bromobenzonitrile.

Preparation 13″:N-(4-{[tert-Butyl(dimethyl)silyl]oxy}phenyl)-3-fluoroaniline

The procedure is in accordance with the process of Preparation 1″,replacing the 4-bromo-1-methyl-1H-pyrazole used in Step B with1-bromo-3-fluorobenzene.

Preparation 14″:N-(4-{[tert-Butyl(dimethyl)silyl]oxy}phenyl)-3,4-difluoroaniline

The procedure is in accordance with the process of Preparation 1″,replacing the 4-bromo-1-methyl-1H-pyrazole used in Step B with4-bromo-1,2-difluorobenzene.

Preparation 15″:4-{[tert-Butyl(dimethyl)silyl]oxy}-N-{4-[(3,3-difluoropiperidin-1-yl)methyl]phenyl}anilineStep A: 1-(4-Bromobenzyl)-3,3-difluoropiperidine

To a solution of 4-bromobenzaldehyde (500 mg; 2.7 mmol) in 12 mL ofdichloromethane there are added, in the order stated,3,3-difluoropiperidine hydrochloride (470 mg; 3 mmol), sodiumtriacetoxyborohydride (860 mg; 4 mmol) and acetic acid (0.17 mL; 3mmol). After stirring for 1 hour at ambient temperature, the reactionmixture is poured over a mixture of aqueous sodium bicarbonate anddichloromethane. The aqueous phase is extracted with dichloromethane.The organic phase is dried over MgSO₄, concentrated to dryness and theresidue is purified by chromatography over silica gel (CH₂Cl₂/MeOHgradient) to provide the expected product in the form of an oil.

¹H NMR: δ (400 MHz; dmso-d6; 300K): 7.55 (dd, 2H); 7.25 (dd, 2H); 3.55(s, 2H); 2.7 (t, 2H); 2.35 (t, 2H); 1.85 (m, 2H); 1.65 (m, 2H)

Step B:4-{[tert-Butyl(dimethyl)silyl]oxy}-N-{4-[(3,3-difluoropiperidin-1-yl)methyl]-phenyl}aniline

The procedure is in accordance with the process of Preparation 1″,replacing the 4-bromo-1-methyl-1H-pyrazole used in Step B with1-[(4-bromophenyl)methyl]-3,3-difluoropiperidine.

Preparation 16″:N-(4-{[tert-Butyl(dimethyl)silyl]oxy}phenyl)quinolin-6-amine

The procedure is in accordance with the process of Preparation 1″,replacing the 4-bromo-1-methyl-1H-pyrazole used in Step B with6-bromo-quinoline.

IR: ν —NH—: 3300 cm⁻¹

Preparation 17″:N-(4-{[tert-Butyl(dimethyl)silyl]oxy}phenyl)-2-methylpyridin-4-amine

The procedure is in accordance with the process of Preparation 1″,replacing the 4-bromo-1-methyl-1H-pyrazole used in Step B with4-bromo-2-methyl-pyridine.

IR: ν —NH—: 3200 and 3100 cm⁻¹

Preparation 18″:N-(4-{[tert-Butyl(dimethyl)silyl]oxy}phenyl)-1-methyl-1H-pyrrolo[2,3-b]pyridin-5-amine

The procedure is in accordance with the process of Preparation 1″,replacing the 4-bromo-1-methyl-1H-pyrazole used in Step B with5-bromo-1-methyl-1H-pyrrolo[2,3-b]pyridine (obtained in accordance witha protocol from the literature: Heterocycles, 60(4), 865, 2003).

IR: ν:—NH—: 3278 cm⁻¹; ν: aromatic —C═C— moieties: 1605 cm⁻¹

Preparation 19″:N-(4-{[tert-Butyl(dimethyl)silyl]oxy}phenyl)pyridin-3-amine

The procedure is in accordance with the process of Preparation 1″,replacing the 4-bromo-1-methyl-1H-pyrazole used in Step B with3-bromo-pyridine.

Preparation 20″:4-{[tert-Butyl(dimethyl)silyl]oxy}-N-{4-[(3,3-difluoropiperidin-1-yl)-ethyl]phenyl}anilineStep A: 2-(4-Bromophenyl)-1-(3,3-difluoropiperidin-1-yl)ethanone

To a solution of 4-bromophenylacetic acid (4 g; 18.6 mmol) and3,3-difluoropiperidine hydrochloride (2.5 g; 20.4 mmol) indichloromethane (190 mL) there are added EDC (3.8 g; 22.3 mmol), HOBt (3g; 22.3 mmol) and triethylamine (1.3 mL; 593 mmol). The reaction mixtureis stirred for 17 hours at ambient temperature and is then poured over amixture of aqueous sodium bicarbonate and ethyl acetate. The aqueousphase is extracted with ethyl acetate. The organic phase is washed with0.1N hydrochloric acid, with water and with brine before being driedover MgSO₄ and concentrated to dryness. The residue is purified bychromatography over silica gel (petroleum ether/ethyl acetate gradient)to provide the expected product in the form of a solid.

¹H NMR: δ (400 MHz; dmso-d6; 300K): 7.5 (d, 2H); 7.2 (d, 2H); 3.8 (t,2H); 3.7 (s, 3H); 3.5 (t, 2H); 2 (m, 2H); 1.6 (m, 2H)

Step B: 1-[2-(4-Bromophenyl)ethyl]-3,3-difluoropiperidine

To a solution of the compound of Step A (4.6 g; 14.5 mmol) in anhydrousTHF (145 mL) there is added a 1M solution of borane dimethyl sulphide inTHF (14.5 mL; 14.5 mmol). The reaction mixture is heated at 80° C. over3 hours and then the solvent is evaporated off under reduced pressure.The residue is treated with methanol (50 mL) and then with 5N HCl (5.8mL). After stirring overnight at ambient temperature and refluxing for 3hours, the pH of the reaction mixture is adjusted to 8 with saturatedsodium bicarbonate solution; the aqueous phase is then extracted withdichloromethane. The organic phase is dried over MgSO₄ and concentratedto dryness, and the residue is purified by chromatography over silicagel (CH₂Cl₂/MeOH gradient) to provide the expected product in the formof an oil.

¹H NMR: δ (400 MHz; dmso-d6; 300K): 7.45 (d, 2H); 7.20 (d, 2H); 2.71 (m,2H); 2.69 (t, 2H); 2.58 (dd, 2H); 2.45 (dd, 2H); 1.86 (m, 2H); 1.63 (m,2H)

Step C:4-{[tert-Butyl(dimethyl)silyl]oxy}-N-{4-[(3,3-difluoropiperidin-1-yl)ethyl]-phenyl}aniline

The procedure is in accordance with the process of Preparation 1″,replacing the 4-bromo-1-methyl-1H-pyrazole used in Step B by thecompound of Step B.

¹H NMR: δ (400 MHz; dmso-d6; 300K): 7.7 (s, 1H); 7.45 (d, 2H); 7.39 (t,2H); 7.31 (t, 1H); 7.0 (m, 4H); 6.9 (d, 2H); 6.81 (d, 2H); 5.05 (s, 2H);2.7 (t, 2H); 2.6 (t, 2H); 2.5 (t, 2H); 2.45 (t, 2H); 1.89 (m, 2H); 1.68(m, 2H)

Preparation 21″:4-{[tert-Butyl(dimethyl)silyl]oxy}-N-{4-[2-(3,3-difluoropyrrolidin-1-yl)ethyl]phenyl}aniline

The procedure is in accordance with the process of Preparation 19″,replacing the 3,3-difluoropiperidine hydrochloride in Step A with3,3-difluoropyrrolidine hydrochloride.

¹H NMR: δ (400 MHz; dmso-d6; 300K): 7.7 (s, 1H); 7.45 (d, 2H); 7.35 (t,2H); 7.34 (t, 1H); 7.05-6.85 (m, 8H); 5.05 (s, 2H); 2.9 (t, 2H);2.75-2.25 (m, 8H)

Preparation 22″:N-(4-{[tert-Butyl(dimethyl)silyl]oxy}phenyl)-2,6-dimethylpyridin-4-amine

The procedure is in accordance with the process of Preparation 1″,replacing the 4-bromo-1-methyl-1H-pyrazole used in Step B with4-bromo-2,6-dimethylpyridine.

IR: ν: —NH—: 3300 and 2700 cm⁻¹; ν:—Si—O—: 900 cm⁻¹; ν: —Si—C—: 823 cm⁻¹

Preparation 23″:N-(4-{[tert-Butyl(dimethyl)silyl]oxy}phenyl)-1-[2-(morpholin-4-yl)-ethyl]-1H-pyrazol-4-amine

The procedure is in accordance with the process of Preparation 2″,replacing the 5-bromoindole used in Step A with 4-bromo-1H-pyrazole.

¹H NMR: δ (400 MHz; dmso-d6; 300K): 7.61 (s, 1H); 7.25 (s, 1H); 7.18 (s,1H); 6.65 (m, 4H); 4.15 (t, 2H); 3.55 (t, 4H); 2.7 (t, 2H); 2.4 (t, 4H);0.95 (s, 9H); 0.15 (s, 6h)

Preparation 24″:N-(4-{[tert-Butyl(dimethyl)silyl]oxy}phenyl)-3-fluoropyridin-4-amine

The procedure is in accordance with the process of Preparation 1″,replacing the 4-bromo-1-methyl-1H-pyrazole used in Step B with4-bromo-3-fluoro-pyridine.

IR: ν —NH—: 3200 and 3000 cm⁻¹; ν —Si—O—: 900 cm⁻¹; ν —Si—C—: 820 cm⁻¹

Preparation 25″:N-(4-{[tert-Butyl(dimethyl)silyl]oxy}phenyl)imidazo[1,2-a]-pyridin-7-amine

The procedure is in accordance with the process of Preparation 1″,replacing the 4-bromo-1-methyl-1H-pyrazole used in Step B with7-bromoimidazo[1,2-a]pyridine (prepared starting from4-bromopyridin-2-amine in accordance with a protocol in the literature:WO 2008124323 A1).

IR: ν —NH—: 3300-3000 cm⁻¹; ν —C═N—: 1652 cm⁻¹; ν —C═C—: 1610 cm⁻¹; ν—Si—C—: 1236 cm⁻¹; ν —Si—O—: 898 cm⁻¹; ν —Si—C—: 828, 774 cm⁻¹

Preparation 26″:N-(4-{[tert-Butyl(dimethyl)silyl]oxy}phenyl)-2-methyl-imidazo[1,2-a]pyridin-7-amine

The procedure is in accordance with the process of Preparation 1″,replacing the 4-bromo-1-methyl-1H-pyrazole used in Step B with7-bromo-2-methyl-imidazo[1,2-a]pyridine (prepared starting from4-bromopyridin-2-amine in accordance with a protocol in the literature:A. J. Helliot et al J. Heterocyclic Chemistry 19, 1437, 1982).

IR: ν —NH—: 3300-3000 cm⁻¹; ν —C═N—: 1652 cm⁻¹; ν —C═C—: 1610 cm⁻¹; ν—Si—C—: 1236 cm⁻¹; ν —Si—O—: 898 cm⁻¹; ν —Si—C—: 828, 774 cm⁻¹

Preparation 27″:N-(4-{[tert-Butyl(dimethyl)silyl]oxy}phenyl)-6-methylpyridin-3-amine

The procedure is in accordance with the process of Preparation 1″,replacing the 4-bromo-1-methyl-1H-pyrazole used in Step B with3-bromo-6-methyl-pyridine.

IR: ν —NH—: 3251 cm⁻¹; ν aromatic —C═C— moieties: 1605 cm⁻¹

Preparation 28″:N-(4-{[tert-Butyl(dimethyl)silyl]oxy}phenyl)-5-fluoropyridin-3-amine

The procedure is in accordance with the process of Preparation 1″,replacing the 4-bromo-1-methyl-1H-pyrazole used in Step B with3-bromo-5-fluoro-pyridine.

IR: ν —NH—: 3400-3000 cm⁻¹; ν —C—F—: 1245 cm⁻¹

Preparation 29″:N-(4-{[tert-Butyl(dimethyl)silyl]oxy}phenyl)-2-methoxypyridin-4-amine

The procedure is in accordance with the process of Preparation 1″,replacing the 4-bromo-1-methyl-1H-pyrazole used in Step B with4-bromo-2-methoxy-pyridine.

IR: ν —NH—: 3200 and 3000 cm⁻¹; ν aromatic —C═C— moieties: 1618, 1601cm⁻¹

Preparation 30″:N-(4-{[tert-Butyl(dimethyl)silyl]oxy}phenyl)-2-(propan-2-yl)pyridin-4-amine

The procedure is in accordance with the process of Preparation 1″,replacing the 4-bromo-1-methyl-1H-pyrazole used in Step B with4-bromo-2-(propan-2-yl)pyridine.

IR: ν —NH—: 3300 and 3100 cm⁻¹

Preparation 31″:N-(4-{[tert-Butyl(dimethyl)silyl]oxy}phenyl)pyrazolo[1,5-a]-pyrimidin-6-amine

The procedure is in accordance with the process of Preparation 1″,replacing the 4-bromo-1-methyl-1H-pyrazole used in Step B with6-bromopyrazolo[1,5-a]pyrimidine.

IR: ν —NH—: 3272 cm⁻¹; ν —C═N—: 1634 cm⁻¹; ν —C═C—: 1616 cm⁻¹

Preparation 32″:N-(4-{[tert-Butyl(dimethyl)silyl]oxy}phenyl)-3,3a-dihydro[1,2,4]triazolo[1,5-a]pyrimidin-6-amine

The procedure is in accordance with the process of Preparation 1″,replacing the 4-bromo-1-methyl-1H-pyrazole used in Step B with6-bromo-3,3a-dihydro[1,2,4]triazolo[1,5-a]-pyrimidine prepared inaccordance with the literature (WO 2011015343) starting from4H-1,2,4-triazol-3-amine and 2-bromopropanedial.

IR: ν —NH—: 3244 cm⁻¹

Preparation 33″:N-(4-{[tert-Butyl(dimethyl)silyl]oxy}phenyl)pyridin-4-amine 1-oxide

The procedure is in accordance with the process of Preparation 1″,replacing the 4-bromo-1-methyl-1H-pyrazole used in Step B with4-bromopyridine 1-oxide prepared in accordance with the literature (WO2009117269) starting from 4-bromopyridine.

IR: ν —NH—: 3246 cm⁻¹; ν aromatic —C═C— moieties: 1618 cm⁻¹

Mass Spectrum:

Empirical formula: C₁₇H₂₄N₂O₂Si

[M]⁺. measured m/z: 316

[M-O]⁺. measured m/z: 300

[M-C₄H₉]⁺. measured m/z: 259

Preparation 34″:N-[4-[tert-Butyl(dimethyl)silyl]oxyphenyl]-1-methyl-pyridin-1-ium-4-aminechloride

The procedure is in accordance with the process of Preparation 1″,replacing the 4-bromo-1-methyl-1H-pyrazole used in Step B with4-bromo-1-methyl-pyridin-1-ium chloride prepared in accordance with theliterature starting from 4-bromopyridine.

Preparation 35″:N-[4-[tert-Butyl(dimethyl)silyl]oxyphenyl]-1-methyl-pyrazolo[3,4-b]pyridin-5-amine

The procedure is in accordance with the process of Preparation 1″,replacing the 4-bromo-1-methyl-1H-pyrazole used in Step B with5-bromo-1-methyl-pyrazolo[3,4-b]pyridine prepared in accordance with theliterature (WO 2006052568).

¹H NMR (400 MHz, dmso-d6) δ ppm: 8.33 (d, 1H), 7.94 (bs, 1H), 7.92 (s,1H), 7.71 (d, 1H), 6.95 (d, 2H), 6.76 (d, 2H), 4.01 (s, 3H), 0.95 (s,9H), 0.17 (s, 6H)

IR (ATR) cm⁻¹: 3290 ν >OH; 1503 ν Ar; 1249 γ —Si—CH₃

Preparation 36″:N-[4-[tert-Butyl(dimethyl)silyl]oxyphenyl]-3-methyl-pyrazolo[1,5-a]pyrimidin-6-amine

The procedure is in accordance with the process of Preparation 1″,replacing the 4-bromo-1-methyl-1H-pyrazole used in Step B with6-bromo-3-methyl-pyrazolo[1,5-a]pyrimidine prepared in accordance withthe literature (WO 2011015343 and WO2011049917).

¹H NMR (400 MHz, dmso-d6) δ ppm: 8.49 (d, 1H), 8.4 (d, 1H), 7.98 (m,1H), 7.87 (s, 1H), 7 (d, 2H), 6.81 (d, 2H), 2.29 (s, 3H), 0.98 (s, 9H),0.2 (s, 6H)

IR (ATR) cm⁻¹: 3257 ν >NH

The amines NHR₃R₄ wherein R₃ and R₄, each independently of the other,represent an aryl or heteroaryl group are obtained in accordance withprocesses described in the literature (Surry D. S. et al., ChemicalScience, 2011, 2, 27-50, Charles M. D. et al., Organic Letters, 2005, 7,3965-3968). The reaction protecting the hydroxy function of the4-anilinophenol described in Preparation 8″ can be applied to varioussecondary amines NHR₃R₄(as defined hereinbefore) having one or morehydroxy functions, when they are available commercially. Alternatively,the secondary amines having at least one hydroxy substituent may besynthesised directly in a protected form, i.e. starting from reagentswhose hydroxy function has been protected beforehand. Among theprotecting groups, tert-butyl(dimethyl)silyloxy and benzyloxy areespecially preferred.

Among the amines NHR₃R₄ having a hydroxy substituent that are used forsynthesising the compounds of the invention there may be mentioned:4-(4-toluidino)phenol, 4-(4-chloroanilino)phenol,4-(3-fluoro-4-methylanilino)phenol,4-[4-(trifluoromethoxy)anilino]-phenol, 4-[4-hydroxyanilino]phenol,{4-[(1-methyl-1H-indol-6-yl)amino]phenyl}-methanol,4-(2,3-dihydro-1H-indol-6-ylamino)phenol,4-[(1-methyl-2,3-dihydro-1H-indol-6-yl)amino]phenol,4-[(1-methyl-1H-indol-6-yl)amino]phenol,4-[(1-methyl-1H-indol-6-yl)amino]cyclohexanol,4-[(1-methyl-1,2,3,4-tetrahydro-6-quinolinyl)amino]phenol,4-[(4-methyl-3,4-dihydro-2H-1,4-benzoxazin-7-yl)amino]phenol,4-[4-(diethylamino)anilino]-phenol,4-(2,3-dihydro-1H-inden-5-ylamino)phenol,4-[(1-methyl-1H-indazol-5-yl)amino]-phenol,4-[(1′-methyl-1′,2′-dihydrospiro[cyclopropane-1,3′-indol]-5′-yl)amino]phenol,4-[(1,3,3-trimethyl-2,3-dihydro-1H-indol-5-yl)amino]phenol,4-[4-methoxy-3-(trifluoro-methyl)anilino]phenol,4-[4-(methylsulphanyl)-3-(trifluoromethyl)anilino]phenol,2-fluoro-4-[(1-methyl-1H-indol-5-yl)amino]phenol,4-[(1-ethyl-1H-indol-5-yl)amino]phenol,4-[(1-ethyl-2,3-dihydro-1H-indol-5-yl)amino]phenol,4-[(1-isopropyl-2,3-dihydro-1H-indol-5-yl)amino]phenol,4-(butylamino)phenol, 3-[(1-methyl-1H-indol-5-yl)amino]-1-propanol,4-[(1-methyl-1H-indol-5-yl)amino]-1-butanol,4-[(3-fluoro-4-methylphenyl)-amino]phenol,4-[(3-chloro-4-methylphenyl)amino]phenol,4-[(4-fluorophenyl)amino]-phenol,4-[(1-methyl-1H-pyrrolo[2,3-b]pyridin-5-yl)amino]phenol,4-[(4-fluorophenyl)-amino]phenol, 4-[(2-fluorophenyl)amino]phenol,4-[(3-fluorophenyl)amino]phenol, 4-[(2,4-difluorophenyl)amino]phenol,4-[(3,4-difluorophenyl)amino]phenol,3-[(4-hydroxy-phenyl)amino]benzonitrile,4-[(3-methoxyphenyl)amino]phenol, 4-[(3,5-difluorophenyl)-amino]phenol,4-[(3-methylphenyl)amino]phenol, 4-[(4-hydroxyphenyl)amino]benzonitrile,4-[(3-chlorophenyl)amino]phenol, 4-(pyrimidin-2-ylamino)phenol,4-[(cyclobutyl-methyl)amino]phenol,2-[(4-hydroxyphenyl)amino]benzonitrile,4-{[(1-methyl-1H-pyrazol-4-yl)methyl]amino}phenol,4-[(cyclopropylmethyl)amino]phenol,4-{[(1-methyl-1H-pyrazol-3-yl)methyl]amino}phenol,4-(but-2-yn-1-ylamino)phenol, 4-(pyrazin-2-yl-amino)phenol,4-(pyridin-2-ylamino)phenol, 4-(pyridazin-3-ylamino)phenol,4-(pyrimidin-5-ylamino)phenol, 4-(pyridin-3-ylamino)phenol,4-[(3,5-difluoro-4-methoxyphenyl)-amino]phenol,4-(pyridin-4-ylamino)phenol, 4-[(3-fluoro-4-methoxyphenyl)amino]phenol,2-(phenylamino)pyrimidin-5-ol,5-[(4-hydroxyphenyl)amino]-2-methoxybenzonitrile,4-{[3-(trifluoromethyl)phenyl]amino}phenol, 4-(methylamino)phenol,4-(ethylamino)phenol and 4-(propan-2-ylamino)phenol.

The hydroxy function(s) of the secondary amines listed above is (are)protected beforehand by a suitable protecting group prior to anycoupling to an acid derivative of the compound of formula (VII) asdefined in the preceding general process.

EXAMPLE 1N-(4-Hydroxyphenyl)-3-(6-{[(3R)-3-methyl-3,4-dihydroisoquinolin-2(1H)-yl]carbonyl}-1,3-benzodioxol-5-yl)-N-(1-methyl-1H-indol-5-yl)-5,6,7,8-tetrahydroindolizine-1-carboxamideStep A: Methyl3-(6-{[(3R)-3-methyl-3,4-dihydroisoquinolin-2(1H)-yl]carbonyl}-1,3-benzodioxol-5-yl)-5,6,7,8-tetrahydroindolizine-1-carboxylate

To a solution of 2 g of the compound of Preparation 1 in 20 mL ofdichloromethane there are added, at ambient temperature, 5.5 mL ofN,N,N-triethylamine (6.96 mmol), the compound of Preparation 1′ (6.96mmol), and then 0.94 g of hydroxybenzotriazole (HOBT) and 1.34 g of1-ethyl-3-(3′-dimethylaminopropyl)-carbodiimide (EDC) (6.96 mmol). Thereaction mixture is then stirred at ambient temperature overnight; it isthen poured onto a solution of ammonium chloride and extracted withethyl acetate. The organic phase is then dried over magnesium sulphate,and then filtered and evaporated to dryness. The crude product therebyobtained is then purified by chromatography over silica gel(heptane/AcOEt gradient) to yield the expected product.

¹H NMR: δ (400 MHz; dmso-d6; 300K): 7.2-6.8 (m, 4H, aromatic Hs, Htetrahydroisoquinoline); 7.10 (s, 1H, aromatic H, benzodioxole); 6.92(s, 1H, aromatic H, benzodioxole); 6.25 (m, 1H, H tetrahydroindolizine);6.10 (s, 2H, aliphatic Hs, OCH₂O); 4.80 (m, 1H, aliphatic H, Htetrahydroisoquinoline); 4.20 (m, 1H, aliphatic H, Htetrahydroisoquinoline); 4.1-3.5 (m, 3H); 3.60 (s, 3H, COOCH₃); 2.90 (m,2H, aliphatic Hs, H tetrahydroindolizine); 2.45 (m, 2H, aliphatic Hs, Htetrahydroisoquinoline); 1.70 (m, 4H, aliphatic Hs, Htetrahydroindolizine); 0.80 (m, 3H, aliphatic Hs, CH₃-THIQ).

IR: ν: >C═O 1694 cm⁻¹ (conjugated ester); ν: >C═O 1624 cm⁻¹ (amide);ν: >C—Ar 772-742 cm⁻¹

Step B: Lithium3-(6-{[(3R)-3-methyl-3,4-dihydroisoquinolin-2(1H)-yl]carbonyl}-1,3-benzodioxol-5-yl)-5,6,7,8-tetrahydroindolizine-1-carboxylate

To a solution containing 8.26 mmol of the compound of Step A in 24 mL ofdioxane there is added a solution of lithium hydroxide (675 mg, 16.1mmol). The batch is placed in a microwave oven at 140 W, 100° C. for aperiod of 2 hours 30 minutes. The reaction mixture is then filtered andevaporated. The solid thereby obtained is dried at 40° C. in an oven inthe presence of P₂O₅.

Step C:N-(4-{[tert-Butyl(dimethyl)silyl]oxy}phenyl)-3-(6-{[(3R)-3-methyl-3,4-dihydroisoquinolin-2(1H)-yl]carbonyl}-1,3-benzodioxol-5-yl)-N-(1-methyl-1H-indol-5-yl)-5,6, 7,8-tetrahydroindolizine-1-carboxamide

To a solution containing 4.73 mmol of the compound of Step B in 47 mL ofdichloromethane there are added, dropwise, 1.2 mL of oxalyl chloride at0° C. The reaction mixture is stirred at ambient temperature for 11hours and is then co-evaporated several times with dichloromethane. Theproduct thereby obtained is suspended in 37 mL of dichloromethane and isthen added to a solution containing 7.1 mmol of the compound obtained inPreparation 2″ in 10 mL of dichloromethane in the presence of 0.6 mL ofpyridine (7.1 mmol). The batch is stirred at ambient temperatureovernight. The reaction mixture is concentrated, purified bychromatography over silica gel (dichloromethane/methanol gradient) toyield the expected product.

Step D:N-(4-Hydroxyphenyl)-3-(6-{[(3R)-3-methyl-3,4-dihydroisoquinolin-2(1H)-yl]-carbonyl}-1,3-benzodioxol-5-yl)-N-(1-methyl-H-indol-5-yl)-5,6,7,8-tetrahydroindolizine-1-carboxamide

To a solution containing 2.3 mmol of the compound obtained in Step C in4 mL of methanol there is added 0.646 g (11.5 mmol) of potassiumhydroxide dissolved in 8 mL of methanol. The batch is stirred at ambienttemperature for 30 minutes. The reaction mixture is then diluted withdichloromethane and washed successively with 1N HCl solution, saturatedNaHCO₃ solution and then saturated NaCl solution until a neutral pH isobtained. The organic phase is then dried over magnesium sulphate,filtered and evaporated. The crude product thereby obtained is purifiedover silica gel (dichloromethane/methanol gradient) and then lyophilisedto provide the expected product.

High-Resolution Mass Spectroscopy (ESI+):

Empirical formula: C₄₂H₃₈CN₄O₅

[M+H]⁺, calculated: 679.2920

[M+H]⁺, measured: 679.2908

EXAMPLE 2N-(4-Hydroxyphenyl)-3-(6-{[(3R)-3-methyl-3,4-dihydroisoquinolin-2(1H)-yl]carbonyl}-1,3-benzodioxol-5-yl)-N-{1-[2-(morpholin-4-yl)ethyl]-1H-indol-5-yl}-5,6,7,8-tetrahydroindolizine-1-carboxamidehydrochloride

The procedure is in accordance with the processes described in Steps A-Dof Example 1 using the appropriate reagents. After the step ofpurification over silica gel (cf. Step D), the solid is then dissolvedin dichloromethane and 2 mL of 1N HCl in ether are added. The entirebatch is stirred for 1 hour and then evaporated to dryness. Thehydrochloride thereby obtained is dissolved in a mixture ofwater/acetonitrile until dissolution is complete and is thenlyophilised.

Elemental Microanalysis: (%, Theoretical:Measured)

% C=69.32:68.93; % H=5.94:5.74; % N=8.6:8.51; % Cl—=4.35:4.6

Unless otherwise mentioned, the compounds of the following Examples aresynthesised in accordance with the process of Example 1 using, in StepA: (i) the appropriate acid obtained according to one of Preparations 1to 18 and (ii) the appropriate tetrahydroisoquinoline compound obtainedaccording to one of Preparations 1′ to 7′ and, in Step C: (iii) thesuitable NHR₃R₄ amine (a non-exhaustive list is proposed in Preparations1″ to 36″).

EXAMPLE 36-(5-Chloro-2-{[(3R)-3-methyl-3,4-dihydroisoquinolin-2(1H)-yl]-carbonyl}phenyl)-N-(4-hydroxyphenyl)-N-(1-methyl-1H-indol-5-yl)-1,2,3,4-tetrahydropyrrolo[1,2-a]pyrazine-8-carboxamidehydrochloride

The procedure is in accordance with the process of Example 1, replacing,on the one hand, the compound of Preparation 1 used in Step A with thecompound of Preparation 2 and, on the other hand, the compound ofPreparation 1″ used in Step C withN-(4-{[tert-butyl(dimethyl)silyl]oxy}phenyl)-1-methyl-1H-indol-5-amine,it being understood that the product thereby obtained is not subjectedto a step of conversion into a salt in the presence of HCl in ether asis described in Step D of Example 1. The compound thereby obtained isdeprotected in the presence of 10 equivalents of trifluoroacetic acid indichloromethane (10 mL/mmol) at ambient temperature overnight. Theproduct is then isolated by concentrating the reaction mixture todryness. Finally, it is subjected to a step of conversion into a salt inthe presence of HCl in ether.

Elemental Microanalysis: (%, Theoretical:Measured)

% C=67.99:65.52; % H=5.28:4.49; % N=9.91:9.24; % Cl=10.03:9.95; %Cl—=5.02:5.45

High-Resolution Mass Spectroscopy (ESI+):

Empirical formula: C₄₀H₃₆ClN₅O₃

[M+H]⁺, calculated: 670.2585

[M+H]⁺, measured: 670.2587

EXAMPLE 43-[5-Chloro-2-(3,4-dihydroisoquinolin-2(1H)-ylcarbonyl)phenyl]-N-(4-hydroxyphenyl)-N-{1-[2-(morpholin-4-yl)ethyl]-1H-indol-5-yl}-5,6,7,8-tetrahydroindolizine-1-carboxamide

Elemental Microanalysis: % Measured (Theoretical)

% C=70.85(71.65); % H=5.39(5.88); % N=9.11(9.28); % Cl=4.48(4.7)

EXAMPLE 53-[5-Chloro-2-(3,4-dihydroisoquinolin-2(1H)-ylcarbonyl)phenyl]-N-(4-hydroxyphenyl)-N-{1-[2-(morpholin-4-yl)ethyl]-2,3-dihydro-1H-indol-5-yl}-5,6,7,8-tetrahydroindolizine-1-carboxamideStep A:N-(4-{[tert-Butyl(dimethyl)silyl]oxy}phenyl)-3-[5-chloro-2-(3,4-dihydroisoquinolin-2(1H)-ylcarbonyl)phenyl]-N—{1-[2-(morpholin-4-yl)ethyl]-2,3-dihydro-1H-indol-5-yl}-5,6,7,8-tetrahydroindolizine-1-carboxamide

The procedure is in accordance with the protocols described in Steps A-Cof Example 1 using the compound of Preparation 3 and1,2,3,4-tetrahydroisoquinoline in Step A, and the compound ofPreparation 4″ in Step C.

Step B:3-[5-Chloro-2-(3,4-dihydroisoquinolin-2(1H)-ylcarbonyl)phenyl]-N-(4-hydroxy-phenyl)-N-{1-[2-(morpholin-4-yl)ethyl]-2,3-dihydro-H-indol-5-yl}-5,6,7,8-tetrahydroindolizine-1-carboxamide

To a solution of 1.3 g (1.45 mmol) of the compound of Step A in 13 mL ofacetic acid there is added, at ambient temperature, sodiumcyanoborohydride (900 mg; 15 mmol). After stirring for 2 hours, thereaction mixture is concentrated to dryness, and then diluted withmethanol (8 mL) and treated with a 1M solution of potassium hydroxide inmethanol (6.3 mL; 6.3 mmol). After 1 hour at ambient temperature, thereaction mixture is concentrated to dryness, and then chromatographedover silica gel (dichloromethane/methanol gradient) and then lyophilisedto provide the expected product in the form of a powder.

Elemental Microanalysis: % Measured (Theoretical)

% C=70.74(71.46); % H=5.74(6.13); % N=9(9.26); % Cl=4.46(4.69)

EXAMPLE 6N-(4-Hydroxyphenyl)-2-methyl-6-(7-{[(3R)-3-methyl-3,4-dihydroisoquinolin-2(1H)-yl]carbonyl}-2,3-dihydro-1,4-benzodioxin-6-yl)-N-(1-methyl-1H-indol-5-yl)-1,2,3,4-tetrahydropyrrolo[1,2-a]pyrazine-8-carboxamidehydrochloride

The procedure is analogous to that described for Example 7, in Step Asubstituting the compound of Preparation 2 with the compound ofPreparation 4.

Elemental Microanalysis: (%, Theoretical:Measured)

% C=69.39:69.13; % H=5.69:4.98; % N=9.41:9.37; % Cl—=4.76:4.65

EXAMPLE 76-(5-Chloro-2-{[(3R)-3-methyl-3,4-dihydroisoquinolin-2(1H)-yl]-carbonyl}phenyl)-N-(4-hydroxyphenyl)-2-methyl-N-(1-methyl-1H-indol-5-yl)-1,2,3,4-tetrahydropyrrolo[1,2-a]pyrazine-8-carboxamideStep A: tert-Butyl8-[(4-{[tert-butyl(dimethyl)silyl]oxy}phenyl)(1-methyl-1H-indol-5-yl)-carbamoyl]-6-(5-chloro-2-{[(3R)-3-methyl-3,4-dihydroisoquinolin-2(1H)-yl]carbonyl}-phenyl)-3,4-dihydropyrrolo[1,2-a]pyrazine-2(1H)-carboxylate

The procedure is in accordance with the protocols described in Steps A-Cof Example 1 using the compounds of Preparations 2 and 1′ in Step A, andthe compound of Preparation 2″ in Step C.

Step B: tert-Butyl6-(5-chloro-2-{[(3R)-3-methyl-3,4-dihydroisoquinolin-2(1H)-yl]-carbonyl}phenyl)-8-[(4-hydroxyphenyl)(1-methyl-1H-indol-5-yl)carbamoyl]-3,4-dihydropyrrolo[1,2-a]pyrazine-2(1H)-carboxylate

To a solution of the compound of Step A (1.1 g; 1.25 mmol) in methanol(6 mL) there is added a 1M solution of potassium hydroxide in methanol(6.2 mL; 6.2 mmol). After 2 hours at ambient temperature, the methanolis evaporated off in vacuo and the residue is taken up in a mixturecomposed of dichloromethane and saturated sodium bicarbonate solution.The combined organic phases are dried over MgSO₄ and concentrated todryness. The residue obtained is purified by chromatography over silicagel (CH₂Cl₂/MeOH gradient) to provide the expected product in the formof a solid.

IR: ν: NH: 3450 cm⁻¹; ν: CO: 1745-1620 cm⁻¹

Step C: tert-Butyl6-(5-chloro-2-{[(3R)-3-methyl-3,4-dihydroisoquinolin-2(1H)-yl]-carbonyl}phenyl)-8-[{4-[(2,2-dimethylpropanoyl)oxy]phenyl}(1-methyl-1H-indol-5-yl)carbamoyl]-3,4-dihydropyrrolo[1,2-a]pyrazine-2(1H)-carboxylate

To a solution of the compound of Step B (0.7 g; 0.93 mmol) indichloromethane (7 mL) there are added, at ambient temperature,triethylamine (0.2 mL; 1.39 mmol) and then pivaloyl chloride (0.11 mL;0.93 mmol). After stirring for 2 hours at ambient temperature, thereaction mixture is washed with water and with brine, dried over MgSO₄and concentrated to dryness. The residue obtained is used as is in thenext Step without being analysed.

Step D: 2,2-Dimethyl4-[{[6-(5-chloro-2-{[(3R)-3-methyl-3,4-dihydroisoquinolin-2(1H)-yl]carbonyl}phenyl)-1,2,3,4-tetrahydropyrrolo[1,2-a]pyrazin-8-yl]carbonyl}(1-methyl-1H-indol-5-yl)amino]phenylpropanoate

To a solution of the compound of the preceding Step (0.82 g; 0.93 mmol)in dichloromethane (9 mL) there is added, at 0° C., trifluoroacetic acid(0.7 mL; 13.9 mmol) dropwise. After stirring for 15 hours at ambienttemperature, saturated sodium bicarbonate solution is slowly added tothe reaction mixture and then the phases are separated. The aqueousphase is extracted with dichloromethane. The combined organic phases aredried over MgSO₄ and concentrated to dryness. The residue obtained ispurified by chromatography over silica gel (CH₂Cl₂/MeOH gradient) toprovide the expected product in the form of a solid.

LC/MS: m/z=[M+H]⁺=754.30

Step E: 2,2-Dimethyl4-[{[6-(5-chloro-2-{[(3R)-3-methyl-3,4-dihydroisoquinolin-2(1H)-yl]carbonyl}phenyl)-2-methyl-1,2,3,4-tetrahydropyrrolo[1,2-a]pyrazin-8-yl]carbonyl}(1-methyl-1H-indol-5-yl)amino]phenylpropanoate

To a solution of the compound of the preceding Step (0.41 g; 0.54 mmol)in dichloromethane (2 mL) there are added, at ambient temperature,formaldehyde (48 μL; 1.74 mmol) and then sodium triacetoxyborohydride(161 mg; 0.76 mmol). After stirring for 2 hours at ambient temperature,the reaction mixture is diluted with dichloromethane and then washedwith saturated sodium bicarbonate solution. The organic phase is driedover MgSO₄ and concentrated to dryness. The residue obtained is purifiedby chromatography over silica gel (CH₂Cl₂/MeOH gradient). The expectedproduct is obtained in the form of a solid.

LC/MS: m/z=[M+H]⁺=768.32

Step F:6-(5-Chloro-2-{[(3R)-3-methyl-3,4-dihydroisoquinolin-2(1H)-yl]carbonyl}-phenyl)-N-(4-hydroxyphenyl)-2-methyl-N-(1-methyl-1H-indol-5-yl)-1,2,3,4-tetrahydropyrrolo[1,2-a]pyrazine-8-carboxamide

To a solution of the compound of the preceding Step (0.25 g; 0.32 mmol)in dioxane (1 mL) there is added a solution of lithium hydroxide (27 mg;0.65 mmol) in water (1 mL).

After stirring for 5 hours at ambient temperature, the reaction mixtureis concentrated and diluted with saturated sodium bicarbonate solution.The aqueous phase is extracted with CH₂Cl₂. The organic phase is driedover MgSO₄ and concentrated to dryness. The residue obtained is purifiedby chromatography over silica gel (CH₂Cl₂/MeOH gradient). The expectedproduct is then obtained in the form of a solid.

Elemental Microanalysis: (%, Theoretical:Measured)

% C=71.97:71.51; % H=5.6:5.25; % N=10.24:10.12

EXAMPLE 83-(5-Chloro-2-{[(3R)-3-methyl-3,4-dihydroisoquinolin-2(1H)-yl]-carbonyl}phenyl)-N-(4-hydroxyphenyl)-N-{1-[2-(morpholin-4-yl)ethyl]-1H-indol-5-yl}indolizine-1-carboxamidehydrochloride

Elemental Microanalysis: (%, Theoretical:Measured)

% C=69:69.16; % H=5.41:4.82; % N=8.75:8.69; % Cl—=4.43:4.13

EXAMPLE 96-(5-Fluoro-2-{[(3R)-3-methyl-3,4-dihydroisoquinolin-2(1H)-yl]carbonyl}-phenyl)-N-(4-fluorophenyl)-N-(4-hydroxyphenyl)-1,2,3,4-tetrahydropyrrolo[1,2-a]-pyrazine-8-carboxamidehydrochloride

The procedure is in accordance with the process of Example 1, replacing,on the one hand, the compound of Preparation 1 used in Step A with thecompound of Preparation 6 and, on the other hand, the compound ofPreparation 1″ used in Step C with the compound of Preparation 5″, itbeing understood that the product thereby obtained is not subjected to astep of conversion into a salt in the presence of HCl in ether as isdescribed in Step D of Example 1. The compound thereby obtained isdeprotected in the presence of 10 equivalents of trifluoroacetic acid indichloromethane (10 mL/mmol) at ambient temperature overnight. Theproduct is then isolated by concentrating the reaction mixture todryness. Finally, it is subjected to a step of conversion into a salt inthe presence of HCl in ether.

Elemental Microanalysis: (%, Theoretical:Measured)

% C=67.83:67.41; % H=5.08:4.61; % N=8.55:8.39; % Cl—=5.41:5.28

EXAMPLE 106-(5-Fluoro-2-{[(3R)-3-methyl-3,4-dihydroisoquinolin-2(1H)-yl]-carbonyl}phenyl)-N-(3-fluoro-4-methylphenyl)-N-(4-hydroxyphenyl)-1,2,3,4-tetrahydropyrrolo[1,2-a]pyrazine-8-carboxamidehydrochloride

The procedure is in accordance with the process of Example 1, replacing,on the one hand, the compound of Preparation 1 used in Step A with thecompound of Preparation 6 and, on the other hand, the compound ofPreparation 1″ used in Step C with the compound of Preparation 6″, itbeing understood that the product thereby obtained is not subjected to astep of conversion into a salt in the presence of HCl in ether as isdescribed in Step D of Example 1. The compound thereby obtained isdeprotected in the presence of 10 equivalents of trifluoroacetic acid indichloromethane (10 mL/mmol) at ambient temperature overnight. Theproduct is then isolated by concentrating the reaction mixture todryness. Finally, it is subjected to a step of conversion into a salt inthe presence of HCl in ether.

Elemental Microanalysis: (%, Theoretical:Measured)

% C=68.21:68.29; % H=5.27:4.91; % N=8.37:8.34; % Cl—=5.3:5.17

EXAMPLE 11N-(4-Hydroxyphenyl)-6-(6-{[(3R)-3-methyl-3,4-dihydroisoquinolin-2(1H)-yl]carbonyl}-1,3-benzodioxol-5-yl)-N-(1-methyl-1H-indazol-5-yl)-1,2,3,4-tetrahydropyrrolo[1,2-a]pyrazine-8-carboxamidehydrochloride

The procedure is in accordance with the process of Example 1, replacing,on the one hand, the compound of Preparation 1 used in Step A with thecompound of Preparation 7 and, on the other hand, the compound ofPreparation 1″ used in Step C withN-(4-{[tert-butyl(dimethyl)silyl]oxy}phenyl)-1-methyl-1H-indazol-5-amine,it being understood that the product thereby obtained is not subjectedto a step of conversion into a salt in the presence of HCl in ether asis described in Step D of Example 1. The compound thereby obtained isdeprotected in the presence of 10 equivalents of trifluoroacetic acid indichloromethane (10 mL/mmol) at ambient temperature overnight. Theproduct is then isolated by concentrating the reaction mixture todryness. Finally, it is subjected to a step of conversion into a salt inthe presence of HCl in ether.

Elemental Microanalysis: (%, Theoretical:Measured)

% H=5.2:4.83; % N=11.72:11.64; % Cl—=4.94:5.34; % C=66.99:66.19

EXAMPLE 126-(5-Chloro-2-{[(3R)-3-methyl-3,4-dihydroisoquinolin-2(1H)-yl]-carbonyl}phenyl)-N-(4-hydroxyphenyl)-N-(1-methyl-1H-indazol-5-yl)-1,2,3,4-tetrahydropyrrolo[1,2-a]pyrazine-8-carboxamidehydrochloride

The procedure is in accordance with the process of Example 1, replacing,on the one hand, the compound of Preparation 1 used in Step A with thecompound of Preparation 2 and, on the other hand, the compound ofPreparation 1″ used in Step C withN-(4-{[tert-butyl(dimethyl)silyl]oxy}phenyl)-1-methyl-1H-indazol-5-amine,it being understood that the product thereby obtained is not subjectedto a step of conversion into a salt in the presence of HCl in ether asis described in Step D of Example 1. The compound thereby obtained isdeprotected in the presence of 10 equivalents of trifluoroacetic acid indichloromethane (10 mL/mmol) at ambient temperature overnight. Theproduct is then isolated by concentrating the reaction mixture todryness. Finally, it is subjected to a step of conversion into a salt inthe presence of HCl in ether.

Elemental Microanalysis: (%, Theoretical:Measured)

% C=66.19:65.83; % H=5.13:4.99; % N=11.88:11.85; % Cl—=5.01:5.36

EXAMPLE 13N-(4-Hydroxyphenyl)-3-(6-{[(3S)-3-[2-(morpholin-4-yl)ethyl]-3,4-dihydroisoquinolin-2(1H)-yl]carbonyl}-1,3-benzodioxol-5-yl)-N-phenyl-5,6,7,8-tetrahydroindolizine-1-carboxamidehydrochloride

Elemental Microanalysis: (%, Theoretical:Measured)

% C=69.42:69.47; % H=5.96:5.58; % N=7.36:7.36; % Cl—=4.66:4.42

EXAMPLE 14N-(4-Hydroxyphenyl)-N-(1-methyl-1H-indazol-5-yl)-3-(6-{[(3S)-3-[2-(morpholin-4-yl)ethyl]-3,4-dihydroisoquinolin-2(1H)-yl]carbonyl}-1,3-benzodioxol-5-yl)-5,6,7,8-tetrahydroindolizine-1-carboxamidehydrochloride

Elemental Microanalysis: (%, Theoretical:Measured)

% C=67.76:67.81; % H=5.81:5.63; % N=10.31:10.13; % Cl—=4.35:4.22

EXAMPLE 157-Amino-N-(4-hydroxyphenyl)-3-(6-{[(3R)-3-methyl-3,4-dihydroisoquinolin-2(1H)-yl]carbonyl}-1,3-benzodioxol-5-yl)-N-phenyl-5,6,7,8-tetrahydroindolizine-1-carboxamidehydrochloride Step A: Methyl3′-(6-{[(3R)-3-methyl-3,4-dihydroisoquinolin-2(1H)-yl]carbonyl}-1,3-benzodioxol-5-yl)-5′,6′-dihydro-8′H-spiro[1,3-dioxolane-2,7′-indolizine]-1′-carboxylate

The procedure is in accordance with the protocol of Step A of Example 1,replacing the compound of Preparation 1 with the compound of Preparation8.

Step B: Methyl3-(6-{[(3R)-3-methyl-3,4-dihydroisoquinolin-2(1H)-yl]carbonyl}-1,3-benzodioxol-5-yl)-7-oxo-5,6,7,8-tetrahydroindolizine-1-carboxylate

4.47 mmol of the compound of Step A dissolved in 75 mL of THF arestirred in the presence of 37 mL of 1M HCl at reflux for 15 hours. 100mL of water and 100 mL of ethyl acetate are added to the reactionmixture. There are then added 4 g of NaHCO₃ (4.7 mmol) in the form of apowder until a basic pH is obtained. The compound is extracted withethyl acetate; the organic phase is dried over MgSO₄, filtered andconcentrated to dryness.

Step C: Methyl7-hydroxy-3-(6-{[(3R)-3-methyl-3,4-dihydroisoquinolin-2(1H)-yl]-carbonyl}-1,3-benzodioxol-5-yl)-5,6, 7,8-tetrahydroindolizine-1-carboxylate

To a solution of 4.47 mmol of the compound obtained in Step B in 30 mLof methanol there are added, in portions, 558 mg (14.75 mmol) of sodiumborohydride. The reaction mixture is stirred for 1 hour at ambienttemperature. 50 mL of 1M HCl are then added and the methanol isevaporated off. The aqueous phase is then neutralised using NaHCO₃ andthen extracted with dichloromethane. The organic phase is successivelywashed with H₂O, dried over MgSO₄, filtered and concentrated to dryness.The oil thereby obtained is purified by flash chromatography(dichloromethane/ethanol-ammonia gradient) to yield the expectedproduct.

Step D: Methyl3-(6-{[(3R)-3-methyl-3,4-dihydroisoquinolin-2(1H)-yl]carbonyl}-1,3-benzodioxol-5-yl)-7-(prop-2-en-1-yloxy)-5,6,7,8-tetrahydroindolizine-1-carboxylate

To a suspension of 331 mg (8.26 mmol) of sodium hydride in 15 mL ofanhydrous THF cooled to 0° C. there are added 4.13 mmol of the compoundobtained in Step C. The resulting suspension is stirred for 15 minutesat 0° C. and then a solution of 790 μL (9.1 mmol) of allyl bromide in 10mL of THF is slowly added (over 15 minutes). The reaction mixture isstirred for 1 hour at 0° C., and then for 15 hours at ambienttemperature. The resulting solution is hydrolysed with saturated aqueousNH₄Cl solution. The compound is extracted with ethyl acetate; theorganic phase is dried over MgSO₄, filtered and concentrated to dryness.The oil thereby obtained is purified by flash chromatography(cyclohexane/ethyl acetate gradient) to yield the expected product.

Step E:N-(4-{[tert-Butyl(dimethyl)silyl]oxy}phenyl)-3-(6-{[(3R)-3-methyl-3,4-dihydroisoquinolin-2(1H)-yl]carbonyl}-1,3-benzodioxol-5-yl)-N-phenyl-7-(prop-2-en-1-yloxy)-5,6,7,8-tetrahydroindolizine-1-carboxamide

The procedure is in accordance with the processes described in Steps Band C of Example 1 using the appropriate reagents.

Step F:N-(4-{[tert-Butyl(dimethyl)silyl]oxy}phenyl)-7-hydroxy-3-(6-{[(3R)-3-methyl-3,4-dihydroisoquinolin-2(H)-yl]carbonyl}-1,3-benzodioxol-5-yl)-N-phenyl-5,6,7,8-tetrahydroindolizine-1-carboxamide

There is then carried out a reaction deprotecting the allyl group in thepresence of 1,3-dimethylpyrimidine-2,4,6(1H,3H,5H)-trione (also calleddimethylbarbiturate) and tetrakis(triphenylphosphine)palladium in amixture of methanol and dichloromethane.

Step G:7-Azido-N-(4-{[tert-butyl(dimethyl)silyl]oxy}phenyl)-3-(6-{[(3R)-3-methyl-3,4-dihydroisoquinolin-2(1H)-yl]carbonyl}-1,3-benzodioxol-5-yl)-N-phenyl-5,6,7,8-tetrahydroindolizine-1-carboxamide

To a solution of the compound of Step F (550 mg; 0.72 mmol) in methylenechloride (6 mL) there are added, at ambient temperature, triethylamine(300 μL; 1.8 mmol) and mesyl chloride (0.14 mL; 1.8 mmol). Afterstirring for 20 minutes, the reaction mixture is concentrated to drynessand then diluted with 10 mL of DMSO. 470 mg of NaN₃ in powder form (7.2mmol) are added thereto. The reaction mixture is left for 20 hours atambient temperature and then for 20 hours at 50° C. It is then pouredonto a mixture of dichloromethane and water. The organic phase is washed3 times with water and then with brine, dried over MgSO₄, and thenconcentrated to dryness to yield the expected product which is used asis in the next Step.

Step H:7-Amino-N-(4-hydroxyphenyl)-3-(6-{[(3R)-3-methyl-3,4-dihydroisoquinolin-2(1H)-yl]carbonyl}-1,3-benzodioxol-5-yl)-N-phenyl-5,6,7,8-tetrahydroindolizine-1-carboxamide hydrochloride

To a solution of 550 mg of the compound of Step G (0.7 mmol) in ethanol(10 mL) there are added, at ambient temperature, 20 mg of Pd/C 10%.After stirring for 15 hours under 1 bar of hydrogen, the reactionmixture is passed through a Whatman filter and concentrated to dryness.After purification by column chromatography over silica gel(dichloromethane/methanol gradient), the solid is then dissolved indichloromethane, and 2 mL of 1N HCl in ether are added. The entire batchis stirred for 1 hour and then evaporated to dryness. The hydrochloridethereby obtained is dissolved in a mixture of water/acetonitrile untildissolution is complete and then lyophilised to yield the expectedcompound in the form of a powder.

Elemental Microanalysis: (%, Theoretical:Measured)

% C=69.17:68.68; % H=5.51:5.09; % N=8.27:8.41; % Cl—=5.24:5.28

EXAMPLE 163-(6-{[(3S)-3-(Hydroxymethyl)-3,4-dihydroisoquinolin-2(1H)-yl]-carbonyl}-1,3-benzodioxol-5-yl)-N-(4-hydroxyphenyl)-N-phenyl-5,6,7,8-tetrahydroindolizine-1-carboxamideStep A: Methyl3-(6-{[(3S)-3-(hydroxymethyl)-3,4-dihydroisoquinolin-2(1H)-yl]-carbonyl}-1,3-benzodioxol-5-yl)-5,6,7,8-tetrahydroindolizine-1-carboxylate

The procedure is in accordance with the process in Step A of Example 1using (3S)-1,2,3,4-tetrahydroisoquinolin-3-ylmethanol.

Step B: Methyl3-(6-{[(3S)-3-[(prop-2-en-1-yloxy)methyl]-3,4-dihydroisoquinolin-2(1H)-yl]carbonyl}-1,3-benzodioxol-5-yl)-5,6, 7,8-tetrahydroindolizine-1-carboxylate

To a suspension of NaH (703 mg; 17.6 mmol) in THF (20 mL) there is addeda solution of 7.8 g of the compound of Step A (16 mmol) dissolved in amixture of THF (50 mL) and DMF (30 mL). After stirring for 1 hour thereis added allyl bromide (1.7 mL; 19 mmol). The reaction mixture isstirred for 48 hours at ambient temperature and is then poured onto amixture of ethyl acetate and water. The organic phase is washed 3 timeswith water, and with saturated LiOH solution, dried over MgSO₄ andconcentrated to dryness. After purification by chromatography oversilica gel (dichloromethane/methanol gradient), the expected product isobtained in the form of a solid.

¹H NMR: δ: (500 MHz; dmso-d6; 300K): 7.2-6.9 (m, 4H); 7.05 (m, 1H); 6.9(m, 1H); 6.45-6.1 (m, 1H); 6.15 (m, 2H); 5.9-5.65 (m, 1H); 5.2-5.0 (m,2H); 5.05-3.8 (m, 1H); 4.85-4.25 (m, 2H); 4.3-3.45 (m, 7H); 3.4-2.4 (m,6H); 1.95-1.45 (m, 4H)

Step C:N-[4-(Benzyloxy)phenyl]-N-phenyl-3-(6-{[(3S)-3-[(prop-2-n-1-yloxy)methyl]-3,4-dihydroisoquinolin-2(1H)-yl]carbonyl}-1,3-benzodioxol-5-yl)-5,6, 7,8-tetrahydroindolizine-1-carboxamide

The procedure is in accordance with the processes of Steps B and C ofExample 1 using 4-(benzyloxy)-N-phenylaniline (cf. Preparation 9″).

Step D:3-(6-{[(3S)-3-(Hydroxymethyl)-3,4-dihydroisoquinolin-2(1H)-yl]carbonyl}-1,3-benzodioxol-5-yl)-N-(4-hydroxyphenyl)-N-phenyl-5,6,7,8-tetrahydroindolizine-1-carboxamide

To a suspension of 5.1 g (6.65 mmol) of the compound of Step C in amixture of dichloromethane (7 mL) and methanol (2 mL) there are addeddimethylbarbituric acid (2.1 g; 13.3 mmol) andtetrakis(triphenylphosphine)palladium(0) (300 mg; 0.3 mmol). Afterstirring for 15 hours at 45° C., the reaction mixture is poured onto amixture of ethyl acetate and water. The organic phase is washed twicewith water, dried over MgSO₄, concentrated to dryness and diluted withmethanol (5 mL). The batch is then stirred for 24 hours under a hydrogenatmosphere in the presence of Pd/C (100 mg). The reaction mixture isthen passed through a Whatman filter, concentrated to dryness, thenchromatographed over silica gel (dichloromethane/methanol gradient) andfinally lyophilised to yield the expected product in the form of apowder.

Elemental Microanalysis: %, Measured (Theoretical)

% C=72.38(73); % H=5.22(5.5); % N=6.59(6.55)

EXAMPLE 17N-{3-Fluoro-4-[2-(morpholin-4-yl)ethoxy]phenyl}-N-(4-hydroxyphenyl)-3-(6-{[(3R)-3-methyl-3,4-dihydroisoquinolin-2(1H)-yl]carbonyl}-1,3-benzodioxol-5-yl)indolizine-1-carboxamidehydrochloride

Elemental Microanalysis: (%, Theoretical:Measured)

% C=67.12:66.79; % H=5.26:4.98; % N=6.96:7.17; % Cl—=4.4:4.77

EXAMPLE 183-[6-(3,4-Dihydroisoquinolin-2(1H)-ylcarbonyl)-1,3-benzodioxol-5-yl]-N-{3-fluoro-4-[2-(morpholin-4-yl)ethoxy]phenyl}-N-(4-hydroxyphenyl)indolizine-1-carboxamidehydrochloride

Elemental Microanalysis: %, Measured (Theoretical)

% C=66.99(66.79); % H=4.93(5.1); % N=7.11(7.08); % Cl—=4.46(4.48)

EXAMPLE 19N-(4-Hydroxyphenyl)-3-(5-methyl-2-{[(3S)-3-[2-(morpholin-4-yl)ethyl]-3,4-dihydroisoquinolin-2(1H)-yl]carbonyl}phenyl)-N-phenyl-5,6,7,8-tetrahydroindolizine-1-carboxamidehydrochloride

Elemental Microanalysis: (%, Theoretical:Measured)

% C=72.26:72.51; % H=6.48:6.13; % N=7.66:7.71; % Cl=4.85:4.95; %Cl—=4.85:4.64

EXAMPLE 20N-(4-Hydroxyphenyl)-3-(2-{[(3S)-3-[2-(morpholin-4-yl)ethyl]-3,4-dihydroisoquinolin-2(1H)-yl]carbonyl}phenyl)-N-phenyl-5,6,7,8-tetrahydroindolizine-1-carboxamidehydrochloride

Elemental Microanalysis: (%, Theoretical:Measured)

% C=72:71.11; % H=6.32:5.94; % N=7.81:7.65; % Cl—=4.94:5.08

EXAMPLE 21N-(4-Hydroxyphenyl)-3-(6-{[(3R)-3-methyl-3,4-dihydroisoquinolin-2(1H)-yl]carbonyl}-1,3-benzodioxol-5-yl)-N-(pyridin-4-yl)indolizine-1-carboxamidehydrochloride

Elemental Microanalysis: (%, Theoretical:Measured)

% C=69.24:69.12; % H=4.74:4.23; % N=8.5:8.45; % Cl—=5.38:5.2

EXAMPLE 22N-(4-Hydroxyphenyl)-6-(6-{[(3S)-3-[2-(morpholin-4-yl)ethyl]-3,4-dihydroisoquinolin-2(1H)-yl]carbonyl}-1,3-benzodioxol-5-yl)-N-phenylpyrrolo[1,2-a]-pyrimidine-8-carboxamidehydrochloride

Elemental Microanalysis: (%, Theoretical:Measured)

% C=68.11:66.66; % H=5.32:4.93; % N=9.24:8.84; % Cl—=4.68:5.78

High-Resolution Mass Spectroscopy (ESI+):

Empirical formula: C₄₃H₃₉N₅O₆

[M+H]⁺, calculated: 655.2915

[M+H]⁺, measured: 655.2915

EXAMPLE 23N-(3-Cyanophenyl)-N-(4-hydroxyphenyl)-3-(6-{[(3S)-3-[2-(morpholin-4-yl)ethyl]-3,4-dihydroisoquinolin-2(1H)-yl]carbonyl}-1,3-benzodioxol-5-yl)-5,6,78-tetrahydroindolizine-1-carboxamidehydrochloride

Elemental Microanalysis: (%, Theoretical:Measured)

% C=68.74:68.59; % H=5.64:5.5; % N=8.91:8.98; % Cl—=4.51:4.48

EXAMPLE 24N-(3-Fluorophenyl)-N-(4-hydroxyphenyl)-3-(6-{[(3S)-3-[2-(morpholin-4-yl)ethyl]-3,4-dihydroisoquinolin-2(1H)-yl]carbonyl}-1,3-benzodioxol-5-yl)-5,6,7,8-tetrahydroindolizine-1-carboxamidehydrochloride

Elemental Microanalysis: (%, Theoretical:Measured)

% C=67.81:67.45; % H=5.69:5.61; % N=7.19:7.42; % Cl—=4.55:4.84

EXAMPLE 25N-(3,4-Difluorophenyl)-N-(4-hydroxyphenyl)-3-(6-{[(3S)-3-[2-(morpholin-4-yl)ethyl]-3,4-dihydroisoquinolin-2(1H)-yl]carbonyl}-1,3-benzodioxol-5-yl)-5,6,7,8-tetrahydroindolizine-1-carboxamidehydrochloride

Elemental Microanalysis: (%, Theoretical:Measured)

% C=66.28:66.56; % H=5.44:5.25; % N=7.03:7.21; % Cl—=4.45:4.32

EXAMPLE 26N-(3-Fluorophenyl)-3-(6-{[(3S)-3-[2-(morpholin-4-yl)ethyl]-3,4-dihydroisoquinolin-2(1H)-yl]carbonyl}-1,3-benzodioxol-5-yl)-N-phenyl-5,6,7,8-tetrahydroindolizine-1-carboxamidehydrochloride

Elemental Microanalysis: (%, Theoretical:Measured)

% C=69.24:70.16; % H=5.81:5.79; % N=7.34:7.47; % Cl—=4.64:4.58

EXAMPLE 273-(5-Chloro-2-{[(3S)-3-[2-(morpholin-4-yl)ethyl]-3,4-dihydroisoquinolin-2(1H)-yl]carbonyl}phenyl)-N-(3-fluorophenyl)-N-(4-hydroxyphenyl)-5,6,7,8-tetrahydroindolizine-1-carboxamidehydrochloride

Elemental Microanalysis: (%, Theoretical:Measured)

% C=67.1:67.68; % H=5.63:5.4; % N=7.28:7.34; % Cl—=4.61:4.59

EXAMPLE 28N-(4-Hydroxyphenyl)-3-(5-methoxy-2{[(3S)-3-[2-(morpholin-4-yl)ethyl]-3,4-dihydroisoquinolin-2(1H)-yl]carbonyl}phenyl)-N-phenyl-5,6,7,8-tetrahydroindolizine-1-carboxamidehydrochloride

Elemental Microanalysis: (%, Theoretical:Measured)

% C=70.72:70.05; % H=6.34:5.95; % N=7.5:7.33; % Cl—=4.74:4.74

EXAMPLE 29N-(4-Hydroxyphenyl)-3-(4-methoxy-2-{[(3S)-3-[2-(morpholin-4-yl)ethyl]-3,4-dihydroisoquinolin-2(1H)-yl]carbonyl}phenyl)-N-phenyl-5,6,7,8-tetrahydroindolizine-1-carboxamidehydrochloride

Elemental Microanalysis: (%, Theoretical:Measured)

% C=70.72:68.96; % H=6.34:5.78; % N=7.5:7.24; % Cl—=4.74:4.62

High-Resolution Mass Spectroscopy (ESI+):

Empirical formula: C₄₄H₄₆N₄O₅

[M+H]⁺, calculated: 711.3546

[M+H]⁺, measured: 711.3540

EXAMPLE 30N-{4-[(3,3-Difluoropiperidin-1-yl)methyl]phenyl}-N-(4-hydroxyphenyl)-3-(6-{[(3R)-3-methyl-3,4-dihydroisoquinolin-2(1H)-yl]carbonyl}-1,3-benzodioxol-5-yl)indolizine-1-carboxamidehydrochloride

Elemental Microanalysis: (%, Theoretical:Measured)

% C=68.31:69.12; % H=5.22:4.93; % N=7.08:6.96; % Cl—=4.48:4.07

EXAMPLE 31N-(4-Hydroxyphenyl)-3-(6-{[(3R)-3-methyl-3,4-dihydroisoquinolin-2(1H)-yl]carbonyl}-1,3-benzodioxol-5-yl)-N-(quinolin-6-yl)indolizine-1-carboxamidehydrochloride

Elemental Microanalysis: (%, Theoretical:Measured)

% C=71.13:71.29; % H=4.69:4.39; % N=7.9:8.14; % Cl—=5:4.5

EXAMPLE 32N-(4-Hydroxyphenyl)-3-(6-{[(3R)-3-methyl-3,4-dihydroisoquinolin-2(1H)-yl]carbonyl}-1,3-benzodioxol-5-yl)-N-(2-methylpyridin-4-yl)indolizine-1-carboxamidehydrochloride

Elemental Microanalysis: (%, Theoretical:Measured)

% C=69.59:69.81; % H=4.94:4.53; % N=8.32:8.59; % Cl—=5.27:5.01

EXAMPLE 33N-(4-Hydroxyphenyl)-3-(6-{[(3R)-3-methyl-3,4-dihydroisoquinolin-2(1H)-yl]carbonyl}-1,3-benzodioxol-5-yl)-N-(1-methyl-1H-pyrrolo[2,3-b]pyridin-5-yl)indolizine-1-carboxamidehydrochloride

Elemental Microanalysis: (%, Theoretical:Measured)

% C=69.14:70.09; % H=4.81:4.55; % N=9.83:10.09; % Cl—=4.98:3.26

EXAMPLE 34N-(4-Hydroxyphenyl)-3(6-{[(3R)-3-methyl-3,4-dihydroisoquinolin-2(1H)-yl]carbonyl}-1,3-benzodioxol-5-yl)-N-(pyridin-3-yl)indolizine-1-carboxamidehydrochloride

Elemental Microanalysis: (%, Theoretical:Measured)

% C=69.24:70.21; % H=4.74:4.42; % N=8.5:8.51; % Cl—=5.38:3.33

EXAMPLE 35N-{4-[2-(3,3-Difluoropiperidin-1-yl)ethyl]phenyl}-N-(4-hydroxyphenyl)-3-(6-{[(3R)-3-methyl-3,4-dihydroisoquinolin-2(1H)-yl]carbonyl}-1,3-benzodioxol-5-yl)indolizine-1-carboxamidehydrochloride

Elemental Microanalysis: (%, Theoretical:Measured)

% C=68.61:67.96; % H=5.38:5.14; % N=6.96:6.76; % Cl—=4.4:4.36

EXAMPLE 36N-{4-[2-(3,3-Difluoropyrrolidin-1-yl)ethyl]phenyl}-N-(4-hydroxyphenyl)-3-(6-{[(3R)-3-methyl-3,4-dihydroisoquinolin-2(1H)-yl]carbonyl}-1,3-benzodioxol-5-yl)indolizine-1-carboxamidehydrochloride

Elemental Microanalysis: (%, Theoretical:Measured)

% C=68.31:68.51; % H=5.22:4.85; % N=7.08:6.83; % Cl—=4.48:4.48

EXAMPLE 373-(6-{[(3S)-3-(2-Aminoethyl)-3,4-dihydroisoquinolin-2(1H)-yl]carbonyl}-1,3-benzodioxol-5-yl)-N-(4-hydroxyphenyl)-N-phenyl-5,6,7,8-tetrahydroindolizine-1-carboxamidehydrochloride Step A: Methyl3-(6-{[(3S)-3-{2-[(tert-butoxycarbonyl)amino]ethyl}-3,4-dihydroisoquinolin-2(1H)-yl]carbonyl}-1,3-benzodioxol-5-yl)-5,6, 7,8-tetrahydroindolizine-1-carboxylate

To a solution of 2 g of the compound of Preparation 1 in 20 mL ofdichloromethane there are added, at ambient temperature, 5.5 mL ofN,N,N-triethylamine (6.96 mmol), the compound of Preparation 3′ (6.96mmol), and then 0.94 g of hydroxybenzotriazole (HOBT) and 1.34 g of1-ethyl-3-(3′-dimethylaminopropyl)-carbodiimide (EDC) (6.96 mmol). Thereaction mixture is then stirred at ambient temperature overnight, andit is then poured onto ammonium chloride solution and extracted withethyl acetate. The organic phase is then dried over magnesium sulphate,and then filtered and evaporated to dryness. The crude product therebyobtained is then purified by chromatography over silica gel(heptane/AcOEt gradient) to yield the expected product.

¹H NMR: δ (400 MHz; dmso-d6; 300K): 7.2-6.8 (m, 4H, aromatic Hs, Htetrahydroisoquinoline); 7.15-6.90 (m, 4H, aromatic H,tetrahydroisoquinoline); 7.00-6.80 (m, 2H, aromatic H, benzodioxole);6.68+6.55+6.25 (m, 1H, NH); 6.50-6.05 (m, 1H, aromatic H,tetrahydroindolizine); 6.12 (m, 2H, aliphatic Hs, OCH₂O); 4.95+4.20+4.10(m, 2H, aliphatic H, CH₂N tetrahydroisoquinoline); 4.85+4.78+3.80 (m,1H, aliphatic H, CH tetrahydroisoquinoline); 4.00-3.40 (m, 2H, aliphaticHs, CH₂N tetrahydroindolizine); 3.70-3.50 (m, 3H, COOCH₃); 2.95-2.45 (m,2H, aliphatic Hs, CH₂NHBoc); 2.98-2.30 (m, 2H, aliphatic Hs, CH₂Ctetrahydroindolizine); 3.00+2.60+2.42 (m, 2H, aliphatic Hs, CH₂CHtetrahydroindolizine); 1.95-1.40 (m, 4H, aliphatic Hs, CH₂CH₂tetrahydroindolizine); 1.35-1.25 (m, 9H, aliphatic Hs, tBu); 1.50-1.15(m, 2H, aliphatic Hs, CH₂CH₂NHBoc)

Step B: Lithium3-(6-{[(3S)-3-{2-[(tert-butoxycarbonyl)amino]ethyl}-3,4-dihydroisoquinolin-2(1H)-yl]carbonyl}-1,3-benzodioxol-5-yl)-5,6,7,8-tetrahydroindolizine-1-carboxylate

To a solution containing 8.26 mmol of the compound of Step A in 24 mL ofdioxane there is added a solution of lithium hydroxide (675 mg, 16.1mmol). The batch is placed in a microwave oven at 140 W, 100° C. for aperiod of 2 hours 30 minutes. The reaction mixture is then filtered andevaporated. The solid thereby obtained is dried at 40° C. in an oven inthe presence of P₂O₅.

Step C: tert-Butyl(2-{(3S)-2-[(6-{1-[(4-{[tert-butyl(dimethyl)silyl]oxy}phenyl)-(phenyl)carbamoyl]-5,6,7,8-tetrahydroindolizin-3-yl}-1,3-benzodioxol-5-yl)carbonyl]-1,2,3,4-tetrahydroisoquinolin-3-yl}ethyl)carbamate

To a solution containing 4.73 mmol of the compound of Step B in 47 mL ofdichloromethane there are added, dropwise, 1.2 mL of oxalyl chloride at0° C. The reaction mixture is stirred at ambient temperature for 11hours and is then co-evaporated several times with dichloromethane. Theproduct thereby obtained is suspended in 37 mL of dichloromethane, andis then added to a solution containing 7.1 mmol of the compound obtainedin Preparation 8″ in 10 mL of dichloromethane in the presence of 0.6 mLof pyridine (7.1 mmol). The batch is stirred at ambient temperatureovernight.

The reaction mixture is concentrated and purified by chromatography oversilica gel (dichloromethane/methanol gradient) to yield the expectedproduct.

¹H NMR: δ (400 MHz; dmso-d6; 300K): 7.0 (m, 11H, aromatic Hs, Ph+4H,tetrahydroisoquinoline+2H, PhO); 6.80-6.65 (m, 2H, aromatic Hs, PhO);6.95-6.85 (m, 2H, aromatic H, benzodioxole); 6.70+6.40 (3tl, 1H, NH);6.10 (m, 2H, aliphatic Hs, OCH₂O); 5.25-4.85 (m, 1H, aromatic H,tetrahydroindolizine); 5.00+4.00 (m, 2H, aliphatic H, CH₂Ntetrahydroisoquinoline); 4.90-3.60 (m, 1H, aliphatic H, CHtetrahydroisoquinoline); 4.10-3.40 (m, 2H, aliphatic Hs, CH₂Ntetrahydroindolizine); 3.00-2.50 (m, 2H, aliphatic Hs, CH₂Ctetrahydroindolizine); 3.00+2.40 (m, 2H, aliphatic Hs, CH₂CHtetrahydroindolizine); 3.00-2.50 (m, 2H, aliphatic Hs, CH₂NHBoc);1.80-1.50 (m, 4H, aliphatic Hs, CH₂CH₂ tetrahydroindolizine); 1.50-1.30(m, 2H, aliphatic Hs, CH₂CH₂NHBoc); 1.35 (2s, 9H, aliphatic Hs, tBu);0.90 (s, 9H, aliphatic Hs, tBu-Si); 0.10 (m, 6H, aliphatic Hs, Me-Si)

Step D:3-(6-{[(3S)-3-(2-Aminoethyl)-3,4-dihydroisoquinolin-2(1H)-yl]carbonyl}-1,3-benzodioxol-5-yl)-N-(4-hydroxyphenyl)-N-phenyl-5,6,7,8-tetrahydroindolizine-1-carboxamidehydrochloride

To a solution of 800 mg (0.92 mmol) of the compound of Step C in 10 mLof methanol there are added 258 mg (4.60 mmol) of KOH. After stirringfor 3 hours at ambient temperature, the reaction mixture is treated with4M HCl solution in 6 mL of dioxane. After stirring for 2 hours atambient temperature, the reaction mixture is concentrated and treatedwith saturated aqueous NaHCO₃ solution and extracted with methylenechloride. The organic phase is then dried over magnesium sulphate, andthen filtered and evaporated to dryness. The crude product therebyobtained is then purified by chromatography over silica gel(dichloromethane/methanol gradient). The compound is then dissolved in 5mL of dichloromethane, and 2.5 mL of 1M HCl in ether are added. Thecompound is filtered off and dried in vacuo. The expected product isobtained in the form of a foam.

Elemental Microanalysis: (%, Theoretical:Measured)

% C=69.51:69.53; % H=5.69:5.27; % N=8.11:8.04; % Cl—=5.13:5.2

High-Resolution Mass Spectroscopy (ESI+):

Empirical formula: C₄₀H₃₈N₄O₅

[M+H]⁺, calculated: 655.2915

[M+H]⁺, measured: 655.2915

¹H NMR: δ (400 MHz; dmso-d6; 300K): 9.55+9.45 (2s, 1H, OH); 7.80+7.75(2s, 3H, NH3⁺); 7.46-6.55 (m, 11H, aromatic Hs, Ph+4H,tetrahydroisoquinoline+2H, PhO); 6.90-6.55 (m, 2H, aromatic Hs, PhO);7.00-6.70 (several s, 2H, aromatic H, benzodioxole); 5.35-5.00 (severals, 1H, aromatic H, tetrahydroindolizine); 6.10 (several s, 2H, aliphaticHs, OCH₂O); 5.00-3.35 (several m, 4H, aliphatic H, CH₂Ntetrahydroisoquinoline+CH₂N tetrahydroindolizine); 4.85+4.75+3.60(several m, 1H, aliphatic H, CH tetrahydroisoquinoline); 2.85-2.45(several m, 2H, aliphatic Hs, CH₂NH₂); 3.00-2.45 (several m, 2H,aliphatic Hs, CH₂C tetrahydroindolizine); 3.05+2.30 (several m, 2H,aliphatic Hs, CH₂CH tetrahydroisoquinoline); 1.85-1.40 (several m, 2H,aliphatic Hs, CH₂ tetrahydroisoquinoline); 1.95-1.35 (several m, 2H,aliphatic Hs, CH₂ tetrahydroisoquinoline); 1.75-1.40 (several m, 2H,aliphatic Hs, CH₂CH₂NH₂)

IR: ν: —OH: 3375 cm⁻¹ (phenol); ν: —NH3⁺: 3500-2300 cm⁻¹ (salt ofprimary amine); ν: >C═O 1612 cm⁻¹+shoulder (amide)

EXAMPLE 38N-(4-Hydroxyphenyl)-3-(6-{[(3R)-3-[3-(morpholin-4-yl)propyl]-3,4-dihydroisoquinolin-2(1H)-yl]carbonyl}-1,3-benzodioxol-5-yl)-N-phenyl-5,6,7,8-tetrahydroindolizine-1-carboxamidehydrochloride

Elemental Microanalysis: (%, Theoretical:Measured)

% C=69.71:69.62; % H=6.11:5.67; % N=7.23:7.12; % Cl—=4.57:4.81

EXAMPLE 39N-(2,6-Dimethylpyridin-4-yl)-N-(4-hydroxyphenyl)-3-(6-{[(3R)-3-methyl-3,4-dihydroisoquinolin-2(1H)-yl]carbonyl}-1,3-benzodioxol-5-yl)indolizine-1-carboxamidehydrochloride

Elemental Microanalysis: (%, Theoretical:Measured)

% C=69.91:69.68; % H=5.13:4.78; % N=8.15:8.03; % Cl—=5.16:5.16

EXAMPLE 40N-(4-Hydroxyphenyl)-3-(6-{[(3R)-3-methyl-3,4-dihydroisoquinolin-2(1H)-yl]carbonyl}-1,3-benzodioxol-5-yl)-N-phenyl-5,6,7,8-tetrahydroindolizine-1-carboxamide

Elemental Microanalysis: (%, Theoretical:Measured)

% C=74.86:74.88; % H=5.64:5.31; % N=6.72:6.78

EXAMPLE 413-(6-{[(3S)-3-[2-(3,3-Difluoropiperidin-1-yl)ethyl]-3,4-dihydroisoquinolin-2(1H)-yl]carbonyl}-1,3-benzodioxol-5-yl)-N-(4-hydroxyphenyl)-N-phenyl-5,6,7,8-tetrahydroindolizine-1-carboxamidehydrochloride

Elemental Microanalysis: (%, Theoretical:Measured)

% C=67.96:68.34; % H=5.7:5.4; % N=7.04:6.97; % Cl—=4.46:4.27

EXAMPLE 42N-(4-Hydroxyphenyl)-3-(6-{[(3R)-3-methyl-3,4-dihydroisoquinolin-2(1H)-yl]carbonyl}-1,3-benzodioxol-5-yl)-N-(pyridin-4-yl)-5,6,7,8-tetrahydroindolizine-1-carboxamidehydrochloride

Elemental Microanalysis: (%, Theoretical:Measured)

% C=68.82:69.46; % H=5.32:4.95; % N=8.45:8.48; % Cl—=5.35:4.6

EXAMPLE 433-(6-{[(3S)-3-{2-[(2,2-Difluoroethyl)amino]ethyl}-3,4-dihydroisoquinolin-2(1H)-yl]carbonyl}-1,3-benzodioxol-5-yl)-N-(4-hydroxyphenyl)-N-phenyl-5,6,7,8-tetrahydroindolizine-1-carboxamideStep A: Ethyl3-(6-{[(3S)-3-{2-[(tert-butoxycarbonyl)amino]ethyl}-3,4-dihydroisoquinolin-2(1H)-yl]carbonyl}-1,3-benzodioxol-5-yl)-5,6,7,8-tetrahydroindolizine-1-carboxylate

The process is analogous to that described in Step A of Example 37.

Step B: Ethyl3-(6-{[(3S)-3-{2-[(tert-butoxycarbonyl)(2,2-difluoroethyl)amino]-ethyl}-3,4-dihydroisoquinolin-2(1H)-yl]carbonyl}-1,3-benzodioxol-5-yl)-5,6,7,8-tetrahydroindolizine-1-carboxylate

To a suspension of 337 mg of NaH (60%) (8.41 mmol) in 13 mL ofdimethylformamide there is added, dropwise, a solution of 1.01 g (1.68mmol) of the compound of Step A in 13 mL of dimethylformamide. Theresulting suspension is stirred at ambient temperature for 15 minutesand there are then added 1.08 g (5.04 mmol) of 2,2-difluoroethyltrifluoromethanesulphonate in 13 mL of dimethylformamide. The batch isstirred at ambient temperature for 2 hours. A solution of 20 mL ofsaturated ammonium chloride is added. The solution is extracted withethyl acetate. The organic phase is then dried over MgSO₄, filtered andconcentrated to dryness. After purification by column chromatographyover silica gel (cyclohexane/ethyl acetate), the expected product isobtained in the form of an oil.

High-Resolution Mass Spectroscopy (ESI+):

Empirical formula: C₃₇H₄₃CN₃O₇

[M+H]⁺, calculated: 680.3142

[M+H]⁺, measured: 680.3145

¹H NMR: δ (400 MHz; dmso-d6; 300K): 7.25-6.90 (m, 4H, aromatic Hs,tetrahydroisoquinoline); 7.10-6.75 (m, 2H, aromatic H, benzodioxole);0.40-6.05 (m, 1H, aromatic H, tetrahydroindolizine); 6.10 (m, 2H,aliphatic Hs, OCH₂O); 6.25-5.90 (m, 1H, aliphatic Hs, CHF₂); 4.95-4.10(m, 2H, aliphatic H, CH₂N tetrahydroisoquinoline); 4.80+3.80 (2m, 1H,aliphatic H, CH tetrahydroisoquinoline); 4.10-4.00 (m, 2H, CH₂Et);4.05-3.40 (m, 2H, aliphatic H, CH₂N tetrahydroindolizine); 3.60-2.60 (m,4H, aliphatic H, CH₂CHF₂+CH₂NBoc); 3.00-2.35 (m, 2H, aliphatic Hs, CH₂Ctetrahydroindolizine); 3.00+2.45 (m, 2H, aliphatic Hs, CH₂CHtetrahydroisoquinoline); 1.95+1.40 (m, 4H, aliphatic Hs, CH₂CH₂tetrahydroindolizine); 1.40 (m, 9H, aliphatic Hs, ^(t)Bu); 1.65-1.20 (m,2H, aliphatic Hs, CH₂CH₂NBoc); 1.18+1.10 (2t, 3H, aliphatic Hs CH₃Et)

Step C: tert-Butyl(2-{(3S)-2-[(6-{1-[(4-{[tert-butyl(dimethyl)silyl]oxy}phenyl)(phenyl)-carbamoyl]-5,6,7,8-tetrahydroindolizin-3-yl}-1,3-benzodioxol-5-yl)carbonyl]-1,2,3,4-tetrahydroisoquinolin-3-yl}ethyl)(2,2-difluoroethyl)carbamate

The process is analogous to that described in Steps B and C of Example37.

¹H NMR: δ (400 MHz; dmso-d6; 300K): 7.30-6.60 (m, 9H, aromatic Hs, 4Htetrahydroisoquinoline+Ph); 6.90-6.70 (m, 2H, aromatic H, benzodioxole);6.80-6.60 (m, 4H, PhO); 6.10 (m, 2H, aliphatic Hs, OCH₂O); 6.20-5.90 (m,1H, aliphatic Hs, CHF₂); 5.50-4.80 (4s, 1H, aromatic H,tetrahydroindolizine); 5.20-4.00 (m, 2H, aliphatic H, CH₂Ntetrahydroisoquinoline); 4.80+4.70+3.50 (3m, 1H, aliphatic H, CHtetrahydroisoquinoline); 4.20-3.40 (m, 2H, aliphatic H, CH₂Ntetrahydroindolizine); 3.60-3.10 (m, 4H, aliphatic H, CH₂CHF₂+CH₂NBoc);3.00+2.60 (m, 2H, aliphatic Hs, CH₂CH tetrahydroisoquinoline); 3.00-2.50(m, 2H, aliphatic Hs, CH₂C tetrahydroindolizine); 1.80+1.50 (m, 4H,aliphatic Hs, CH₂CH₂ tetrahydroindolizine); 1.60-1.30 (m, 2H, aliphaticHs, CH₂CH₂NBoc); 1.40-1.30 (m, 9H, aliphatic Hs, tBu); 0.90 (4s, 9H,aliphatic Hs, tBu-Si); 0.10 (4s, 6H, aliphatic Hs, Me-Si)

Step D:3-(6-{[(3S)-3-{2-[(2,2-Difluoroethyl)amino]ethyl}-3,4-dihydroisoquinolin-2(1H)-yl]carbonyl}-1,3-benzodioxol-5-yl)-N-(4-hydroxyphenyl)-N-phenyl-5,6, 7,8-tetrahydroindolizine-1-carboxamide

To a solution of 933 mg (1.00 mmol) of the compound of Step C in 10 mLof methanol there are added 280 mg (5.00 mmol) of KOH. After stirringfor 3 hours at ambient temperature, the reaction mixture is treated with4M HCl solution in 6 mL of dioxane. After stirring for 2 hours atambient temperature, the reaction mixture is concentrated and treatedwith aqueous saturated NaHCO₃ solution and then extracted with methylenechloride. The organic phase is then dried over magnesium sulphate, andthen filtered and evaporated to dryness. The crude product therebyobtained is then purified by chromatography over silica gel(dichloromethane/methanol gradient) to yield the expected product in theform of a foam.

Elemental Microanalysis: (%, Theoretical:Measured)

% C=70.18:69.79; % H=5.61:5.67; % N=7.79:7.7

High-Resolution Mass Spectroscopy (ESI+):

Empirical formula: C₄₂H₄₀F₂N₄O₅

[M+H]⁺, calculated: 655.2915

[M+H]⁺, measured: 655.2915

EXAMPLE 44N-(4-Hydroxyphenyl)-3-(6-{[(3S)-3-[2-(3-methoxyazetidin-1-yl)ethyl]-3,4-dihydroisoquinolin-2(1H)-yl]carbonyl}-1,3-benzodioxol-5-yl)-N-phenyl-5,6,7,8-tetrahydroindolizine-1-carboxamide

Elemental Microanalysis: (%, Theoretical:Measured)

% C=72.91:72.73; % H=6.12:5.67; % N=7.73:7.74

EXAMPLE 45N-(4-Hydroxyphenyl)-3-(6-{[(3R)-3-methyl-3,4-dihydroisoquinolin-2(1H)-yl]carbonyl}-1,3-benzodioxol-5-yl)-N-{t-[2-(morpholin-4-yl)ethyl]-1H-pyrazol-4-yl}indolizine-1-carboxamidehydrochloride

Elemental Microanalysis: (%, Theoretical:Measured)

% C=66.27:66.05; % H=5.43:5.27; % N=11.04:11.07; % Cl—=4.66:4.61

EXAMPLE 46N-(3-Fluoropyridin-4-yl)-N-(4-hydroxyphenyl)-3-(6-{[(3R)-3-methyl-3,4-dihydroisoquinolin-2(1H)-yl]carbonyl}-1,3-benzodioxol-5-yl)indolizine-1-carboxamidehydrochloride

High-resolution mass spectroscopy (ESI+):

Empirical formula: C₃₈H₂₉FN₄O₅

[M+H]⁺, calculated: 641.2195

[M+H]⁺, measured: 641.2195

EXAMPLE 473-(5-Chloro-2-{[(3R)-3-methyl-3,4-dihydroisoquinolin-2(1H)-yl]-carbonyl}phenyl)-N-(4-hydroxyphenyl)-N-(1-methyl-1H-pyrazol-4-yl)-5,6,7,8-tetrahydroindolizine-1-carboxamide

Elemental Microanalysis: (%, Theoretical:Measured)

% C=69.72:69.53; % H=5.53:5.6; % N=11.29:10.85

EXAMPLE 48N-(4-Hydroxyphenyl)-3-(7-{[(3R)-3-methyl-3,4-dihydroisoquinolin-2(1H)-yl]carbonyl}-2,3-dihydro-1,4-benzodioxin-6-yl)-N-(1-methyl-1H-pyrazol-4-yl)-5,6,7,8-tetrahydroindolizine-1-carboxamide

Elemental Microanalysis: (%, Theoretical:Measured)

% C=70.9:70.89; % H=5.79:5.56; % N=10.88:10.8

EXAMPLE 493-(5-Chloro-2-{[(3R)-3-methyl-3,4-dihydroisoquinolin-2(1H)-yl]-carbonyl}phenyl)-N-(4-hydroxyphenyl)-N-(pyridin-4-yl)indolizine-1-carboxamidehydrochloride

Elemental Microanalysis: (%, Theoretical:Measured)

% C=68.42:68.17; % H=4.65:4.48; % N=8.63:8.48; % Cl—=5.46:5.13

EXAMPLE 503-(5-Chloro-2-{[(3R)-3-methyl-3,4-dihydroisoquinolin-2(1H)-yl]-carbonyl}phenyl)-N-(4-hydroxyphenyl)-N-(1-methyl-1H-pyrrolo[2,3-b]pyridin-5-yl)indolizine-1-carboxamide

Elemental Microanalysis: (%, Theoretical:Measured)

% C=72.12:71.58; % H=4.84:4.84; % N=10.51:10.48

EXAMPLE 51N-(4-Hydroxyphenyl)-N-(imidazo[1,2-a]pyridin-7-yl)-3-(6-{[(3R)-3-methyl-3,4-dihydroisoquinolin-2(1H)-yl]carbonyl}-1,3-benzodioxol-5-yl)indolizine-1-carboxamidehydrochloride

Elemental Microanalysis: (%, Theoretical:Measured)

% C=68.81:68.28; % H=4.62:4.59; % N=10.03:9.66; % Cl—=5.08:4.81

EXAMPLE 52N-(4-Hydroxyphenyl)-3-(2-{[(3R)-3-methyl-3,4-dihydroisoquinolin-2(1H)-yl]carbonyl}phenyl)-N-(1-methyl-1H-pyrrolo[2,3-b]pyridin-5-yl)indolizine-1-carboxamide

Elemental Microanalysis: (%, Theoretical:Measured)

% C=76.05:75.88; % H=5.26:5.24; % N=11.09:11.09

EXAMPLE 53N-(4-Hydroxyphenyl)-3-(6-{[(3R)-3-methyl-3,4-dihydroisoquinolin-2(1H)-yl]carbonyl}-1,3-benzodioxol-5-yl)-N-(2-methylimidazo[1,2-a]pyridin-7-yl)indolizine-1-carboxamidehydrochloride

Elemental Microanalysis: (%, Theoretical:Measured)

% C=69.14:69.65; % H=4.81:4.75; % N=9.83:9.79; % Cl—=4.98:4.7

EXAMPLE 54N-(4-Hydroxyphenyl)-3-(6-{[(3R)-3-methyl-3,4-dihydroisoquinolin-2(1H)-yl]carbonyl}-1,3-benzodioxol-5-yl)-N-(6-methylpyridin-3-yl)indolizine-1-carboxamidehydrochloride

Elemental Microanalysis: (%, Theoretical:Measured)

% C=69.59:68.78; % H=4.94:5; % N=8.32:8.33; % Cl—=5.27:5.18

EXAMPLE 55N-(5-Fluoropyridin-3-yl)-N-(4-hydroxyphenyl)-3-(6-{[(3R)-3-methyl-3,4-dihydroisoquinolin-2(1H)-yl]carbonyl}-1,3-benzodioxol-5-yl)indolizine-1-carboxamide

Elemental Microanalysis: (%, Theoretical:Measured)

% C=71.24:70.77; % H=4.56:4.36; % N=8.75:8.82

EXAMPLE 56N-(4-Hydroxyphenyl)-N-(2-methoxypyridin-4-yl)-3-(6-{[(3R)-3-methyl-3,4-dihydroisoquinolin-2(1H)-yl]carbonyl}-1,3-benzodioxol-5-yl)indolizine-1-carboxamide

High-resolution mass spectroscopy (ESI+):

Empirical formula: C₃₉H₃₂N₄O₆

[M+H]⁺, calculated: 653.2395

[M+H]⁺, measured: 653.2385

EXAMPLE 573-[6-(3,4-Dihydroisoquinolin-2(1H)-ylcarbonyl)-1,3-benzodioxol-5-yl]-N-(4-hydroxyphenyl)-N-phenyl-5,6,7,8-tetrahydroindolizine-1-carboxamide

Elemental Microanalysis: %, Measured (Theoretical)

% C=74.17(74.62); % H=5.43(5.44); % N=6.87(6.87)

EXAMPLE 58N-(4-Hydroxyphenyl)-3-(6-{[(3R)-3-methyl-3,4-dihydroisoquinolin-2(1H)-yl]carbonyl}-1,3-benzodioxol-5-yl)-N-[2-(propan-2-yl)pyridin-4-yl]indolizine-1-carboxamidehydrochloride

Elemental Microanalysis: (%, Theoretical:Measured)

% C=70.23:69.95; % H=5.32:5.4; % N=7.99:7.99; % Cl—=5.06:4.92

EXAMPLE 59N-(4-Hydroxyphenyl)-3-(6-{[(3R)-3-methyl-3,4-dihydroisoquinolin-2(1H)-yl]carbonyl}-1,3-benzodioxol-5-yl)-N-(pyrazolo[1,5-a]pyrimidin-6-yl)indolizine-1-carboxamide

Elemental Microanalysis: (%, Theoretical:Measured)

% C=70.68:70.47; % H=4.56:4.61; % N=12.68:12.45

EXAMPLE 603-(5-Fluoro-2-{[(3R)-3-methyl-3,4-dihydroisoquinolin-2(1H)-yl]-carbonyl}phenyl)-N-(4-hydroxyphenyl)-N-(1-methyl-1H-pyrazol-4-yl)indolizine-1-carboxamide

Elemental Microanalysis: %, Measured (Theoretical)

% C=71.85(72.11); % H=4.78(5.04); % N=10.79(11.68)

EXAMPLE 613-(5-Fluoro-2-{[(3R)-3-methyl-3,4-dihydroisoquinolin-2(1H)-yl]-carbonyl}phenyl)-N-(4-hydroxyphenyl)-N-(1-methyl-1H-pyrazol-4-yl)-5,6,7,8-tetrahydroindolizine-1-carboxamide

Elemental Microanalysis: %, Measured (Theoretical)

% C=72.31(71.62); % H=5.6(5.68); % N=10.94(11.6)

EXAMPLE 623-(5-Fluoro-2-{[(3R)-3-methyl-3,4-dihydroisoquinolin-2(1H)-yl]-carbonyl}phenyl)-N-(4-hydroxyphenyl)-N-(pyridin-4-yl)indolizine-1-carboxamide

Elemental Microanalysis: %, Measured (Theoretical)

% C=74.08(74.48); % H=4.82(4.9); % N=8.59(9.39)

EXAMPLE 633-(5-Fluoro-2-{[(3R)-3-methyl-3,4-dihydroisoquinolin-2(1H)-yl]-carbonyl}phenyl)-N-(4-hydroxyphenyl)-N-(1-methyl-1H-pyrrolo[2,3-b]pyridin-5-yl)indolizine-1-carboxamide

Elemental Microanalysis: %, Measured (Theoretical)

% C=73.14(73.95); % H=4.83(4.96); % N=10.29(10.78)

EXAMPLE 64 3-(5-Fluoro-2-{[(3R)-3-rethyl-3,4-dihydroisoquinolin-2(1H)-yl]-carbonyl}phenyl)-N-(4-hydroxyphenyl)-N-(pyridin-4-yl)-5,6,7,8-tetrahydroindolizine-1-carboxamide

Elemental Microanalysis: %, Measured (Theoretical)

% C=74.61(73.98); % H=5.26(5.54); % N=8.94(9.33)

EXAMPLE 653-(5-Fluoro-2-{[(3R)-3-methyl-3,4-dihydroisoquinolin-2(1H)-yl]-carbonyl}phenyl)-N-(4-hydroxyphenyl)-N-(1-methyl-1H-pyrrolo[2,3-b]pyridin-5-yl)-5,6,7,8-tetrahydroindolizine-1-carboxamide

Elemental Microanalysis: %, Measured (Theoretical)

% C=73.59(73.49); % H=5.22(5.55); % N=9.93(10.71)

EXAMPLE 66N-(4-Hydroxyphenyl)-3-(6-{[(3R)-3-methyl-3,4-dihydroisoquinolin-2(1H)-yl]carbonyl}-1,3-benzodioxol-5-yl)-N-([1,2,4]triazolo[1,5-a]pyrimidin-6-yl)-indolizine-1-carboxamide

Elemental Microanalysis: %, Measured (Theoretical)

% C=68.57(68.77); % H=3.92(4.4); % N=14.21(14.77)

High-resolution mass spectroscopy (ESI+):

Empirical formula: C₃₈H₂₉N₇O₅

[M+H]⁺, calculated: 664.2303

[M+H]⁺, measured: 664.2310

EXAMPLE 67N-(4-Hydroxyphenyl)-3-(6-{[(3R)-3-methyl-3,4-dihydroisoquinolin-2(1H)-yl]carbonyl}-1,3-benzodioxol-5-yl)-N-(1-oxidopyridin-4-yl)indolizine-1-carboxamide

Elemental Microanalysis: %, Measured (Theoretical)

% C=69.7(71.46); %% H=4.43(4.73); % N=8.54(8.77)

High-Resolution Mass Spectroscopy (ESI+):

Empirical formula: C₃₈H₃₀N₄O₆

[M+H]⁺, calculated: 639.2238

[M+H]⁺, measured: 639.2234

EXAMPLE 68N-(4-Hydroxyphenyl)-3-(2-{[(3R)-3-methyl-3,4-dihydroisoquinolin-2(1H)-yl]carbonyl}phenyl)-N-(pyridin-4-yl)indolizine-1-carboxamidehydrochloride

Elemental Microanalysis: %, Measured (Theoretical)

% C=71.97(72.25); % H=5.21(5.08); % N=8.99(9.11); % Cl—=5.32(5.76)

High-Resolution Mass Spectroscopy (ESI+):

Empirical formula: C₃₇H₃₀N₄O₃

[M+H]⁺, calculated: 579.2391

[M+H]⁺, measured: 579.2403

EXAMPLE 69N-(4-Hydroxyphenyl)-N-(1-methyl-1H-pyrrolo[2,3-b]pyridin-5-yl)-3-(6-{[(3R)-3-[3-(morpholin-4-yl)propyl]-3,4-dihydroisoquinolin-2(1H)-yl]carbonyl}-1,3-benzodioxol-5-yl)-5,6,7,8-tetrahydroindolizine-1-carboxamidehydrochloride

Elemental Microanalysis: %, Measured (Theoretical)

% C=67.63(68.06); % H=5.27(5.95); % N=10.08(10.13); % Cl—=4.53(4.27)

High-Resolution Mass Spectroscopy (ESI+):

Empirical formula: C₄₇H₄₈N₆O₆

[M+H]⁺, calculated: 793.3708

[M+H]⁺, measured: 793.3704

EXAMPLE 70N-(4-Hydroxyphenyl)-3-(6-{[(3R)-3-methyl-3,4-dihydroisoquinolin-2(1H)-yl]carbonyl}-1,3-benzodioxol-5-yl)-N-(1-methyl-1H-pyrazolo[3,4-b]pyridin-5-yl)indolizine-1-carboxamideStep A:N-[4-[tert-Butyl(dimethyl)silyl]oxyphenyl]-3-[6-[(3R)-3-methyl-3,4-dihydro-1H-isoquinoline-2-carbonyl]-1,3-benzodioxol-5-yl]-N-(1-methyl-1H-pyrazolo[3,4-b]pyridin-5-yl)indolizine-1-carboxamide

The title product is obtained in accordance with the process of Step Aof Example 86, replacing the compound of Preparation 36″ with that ofPreparation 35″.

LCMS: [M+H]⁺=791.4 vs. 791.3 calculated

Step B:N-(4-Hydroxyphenyl)-3-(6-{[(3R)-3-methyl-3,4-dihydroisoquinolin-2(1H)-yl]-carbonyl}-1,3-benzodioxol-5-yl)-N-(1-methyl-1H-pyrazolo[3,4-b]pyridin-5-yl)indolizine-1-carboxamide

The procedure is in accordance with a protocol analogous to thatdescribed in Step D of Example 1. The product thereby obtained issubjected to a step of conversion into a salt in the presence of HCl inether.

IR (ATR) cm⁻¹: 2500 to 3000 ν —OH, 1614 ν >C═O amides, 1236 ν >C—O—C<,740 γ >CH—Ar

Elemental Microanalysis: %, Measured (Theoretical)

% C=71.07(70.99); % H=4.45(4.77); % N=12.37(12.42)

High-Resolution Mass Spectroscopy (ESI+):

Empirical formula: C₄₀H₃₂N₆O₅

[M+H]⁺, calculated: 677.2507

[M+H]⁺, measured: 677.2510

EXAMPLE 714-[(4-Hydroxyphenyl){[3-(6-{[(3R)-3-methyl-3,4-dihydroisoquinolin-2(1H)-yl]carbonyl}-1,3-benzodioxol-5-yl)indolizin-1-yl]carbonyl}amino]-1-methyl-pyridiniumchloride Step A:4-[(4-Hydroxyphenyl){[3-(6-{[(3R)-3-methyl-3,4-dihydroisoquinolin-2(1H)-yl]-carbonyl}-1,3-benzodioxol-5-yl)indolizin-1-yl]carbonyl}amino]-1-methylpyridinium iodide

The compound of Example 21 (311 mg, 0.5 mmol) is dissolved indichloromethane and washed with saturated aqueous sodium hydrogencarbonate solution. After drying the organic phase over magnesiumsulphate and evaporating to dryness, the residue is dissolved in ethanol(30 mL). Methyl iodide (45 μL, 0.7 mmol) is then added and the reactionmixture is heated to 40° C. The solution thereby obtained is evaporatedto dryness. The crude reaction product is purified over a silica gelcolumn using dichloromethane and methanol as solvents. The compound isobtained in the form of a white powder which is used directly in thenext Step.

¹H NMR (500 MHz, dmso-d6) δ ppm: 9.95 (bs, 1H), 8.6-8.45 (m, 2H),8.35-8.05 (several m, 1H), 8.3-8 (several m, 1H), 7.45-6.7 (several m,8H), 7.4-6.9 (several m, 4H), 6.45-6.3 (several s, 1H), 6.45-6.3 (m,2H), 6.15 (s, 2H), 5.05-3.55 (several d, 2H), 4.75/3.8 (m+m, 1H), 4.15(2*s, 3H), 2.95-2.1 (several m, 2H), 1-0.15 (several m, 3H)

Step B:4-[(4-Hydroxyphenyl){[3-(6-{[(3R)-3-methyl-3,4-dihydroisoquinolin-2(1H)-yl]-carbonyl}-1,3-benzodioxol-5-yl)indolizin-1-yl]carbonyl}amino]-1-methylpyridinium chloride

The compound of the preceding Step (320 mg, 0.42 mmol) is dissolved inmethanol (20 mL), and then silver carbonate (173 mg, 0.628 mmol) isadded, in portions, over 10 minutes. The resulting suspension is stirredfor 1 hour at ambient temperature; the precipitate is filtered off andwashed with methanol. The filtrate is concentrated to dryness, and thentreated with 50 mL of 2N hydrochloric acid solution, heated at 60° C.for 30 minutes and then evaporated to dryness. The final product isobtained after purification over a silica C18 column using a 0.1%hydrochloric acid solution and acetonitrile as solvents. The titlecompound is obtained in the form of a white powder which is lyophilisedin a mixture of water/acetonitrile.

IR (ATR) cm⁻¹: 3388 ν —OH phenol, 1650+1627 ν >C═O amides

High-Resolution Mass Spectroscopy (ESI+):

Empirical formula: C₃₉H₃₃N₄O₅

[M]⁺, calculated=637.2445.

[M]⁺, measured=637.2431

The compounds of Examples 72, 73, 77, 78-80, 84 and 85 are synthesisedin accordance with the process of Example 3 using the acid ofPreparation 7, the appropriate 1,2,3,4-tetrahydroisoquinoline or theappropriate compound obtained in accordance with one of Preparations 1′to 7′, and the suitable NHR₃R₄ amine.

EXAMPLE 72N-(4-Hydroxyphenyl)-N-methyl-6-(6-{[(3R)-3-methyl-3,4-dihydroisoquinolin-2(1H)-yl]carbonyl}-1,3-benzodioxol-5-yl)-1,2,3,4-tetrahydropyrrolo[1,2-a]-pyrazine-8-carboxamide

LC/MS (C₃₃H₃₂N₄O₅) 565 [M+H]⁺; RT 1.47 (Method B), it being understoodthat RT denotes retention time

EXAMPLE 73N-Ethyl-N-(4-hydroxyphenyl)-6-(6-{[(3R)-3-methyl-3,4-dihydroisoquinolin-2(1H)-yl]carbonyl}-1,3-benzodioxol-5-yl)-1,2,3,4-tetrahydropyrrolo[1,2-a]-pyrazine-8-carboxamide

LC/MS (C₃₄H₃₄N₄₀₅) 579 [M+H]⁺; RT 1.55 (Method B)

EXAMPLE 743-[6-(3,4-Dihydroisoquinolin-2(1H)-ylcarbonyl)-1,3-benzodioxol-5-yl]-N-(4-hydroxyphenyl)-N-methyl-5,6,7,8-tetrahydroindolizine-1-carboxamide

LC/MS (C₃₃H₃₁N₃₀₅) 550 [M+H]⁺; RT 1.24 (Method B)

EXAMPLE 753-[6-(3,4-Dihydroisoquinolin-2(1H)-ylcarbonyl)-1,3-benzodioxol-5-yl]-N-ethyl-N-(4-hydroxyphenyl)-5,6,7,8-tetrahydroindolizine-1-carboxamide

LC/MS (C₃₄H₃₃N₃O₅) 564 [M+H]⁺; RT 1.30 (Method B)

EXAMPLE 76N-Butyl-3-[6-(3,4-dihydroisoquinolin-2(1H)-ylcarbonyl)-1,3-benzodioxol-5-yl]-N-(4-hydroxyphenyl)-5,6,7,8-tetrahydroindolizine-1-carboxamide

LC/MS (C₃₆H₃₇N₃O₅) 592 [M+H]⁺; RT 1.39 (Method B)

EXAMPLE 77N-Ethyl-N-(4-hydroxyphenyl)-6-(6-{[(3S)-3-methyl-3,4-dihydroisoquinolin-2(1H)-yl]carbonyl}-1,3-benzodioxol-5-yl)-1,2,3,4-tetrahydropyrrolo[1,2-a]-pyrazine-8-carboxamide

LC/MS (C₃₄H₃₄N₄O₅) 579 [M+H]⁺; RT 1.50 (Method B)

EXAMPLE 78N,N-Dibutyl-6-(6-{[(3R)-3-methyl-3,4-dihydroisoquinolin-2(1H)-yl]-carbonyl}-1,3-benzodioxol-5-yl)-1,2,3,4-tetrahydropyrrolo[1,2-a]pyrazine-8-carboxamide

LC/MS (C₃₄H₄₂N₄O₄) 571 [M+H]⁺; RT 1.79 (Method B)

EXAMPLE 79N-Butyl-N-(4-hydroxyphenyl)-6-(6-{[(3R)-3-methyl-3,4-dihydroisoquinolin-2(1H)-yl]carbonyl}-1,3-benzodioxol-5-yl)-1,2,3,4-tetrahydropyrrolo[1,2-a]-pyrazine-8-carboxamide

LC/MS (C₃₆H₃₈N₄O₅) 607 [M+H]⁺; RT 1.65 (Method B)

EXAMPLE 80N-(4-Hydroxyphenyl)-6-(6-{[(3R)-3-methyl-3,4-dihydroisoquinolin-2(1H)-yl]carbonyl}-1,3-benzodioxol-5-yl)-N-(propan-2-yl)-1,2,3,4-tetrahydropyrrolo[1,2-a]pyrazine-8-carboxamide

LC/MS (C₃₅H₃₆N₄O₅) 593 [M+H]⁺; RT 1.58 (Method B)

EXAMPLE 81N-(4-Hydroxyphenyl)-N-methyl-3-(6-{[(3R)-3-methyl-3,4-dihydroisoquinolin-2(1H)-yl]carbonyl}-1,3-benzodioxol-5-yl)-5,6,7,-tetrahydroindolizine-1-carboxamide

LC/MS (C₃₄H₃₃N₃O₅) 564 [M+H]⁺; RT 2.48 (Method A)

EXAMPLE 82N-(4-Hydroxyphenyl)-N-methyl-3-(6-{[(3S)-3-methyl-3,4-dihydroisoquinolin-2(1H)-yl]carbonyl}-1,3-benzodioxol-5-yl)-5,6,7,8-tetrahydroindolizine-1-carboxamide

LC/MS (C₃₄H₃₃N₃O₅) 564 [M+H]⁺; RT 2.55 (Method A)

EXAMPLE 833-[6-(3,4-Dihydroisoquinolin-2(1H)-ylcarbonyl)-1,3-benzodioxol-5-yl]-N-(4-hydroxyphenyl)-N-methylindolizine-1-carboxamide

LC/MS (C₃₃H₂₇N₃O₅) 546 [M+H]⁺; RT 2.40 (Method A)

EXAMPLE 846-[6-(3,4-Dihydroisoquinolin-2(1H)-ylcarbonyl)-1,3-benzodioxol-5-yl]-N-(4-hydroxyphenyl)-N-methyl-1,2,3,4-tetrahydropyrrolo[1,2-a]pyrazine-8-carboxamide

LC/MS (C₃₂H₃₀N₄O₅) 551 [M+H]⁺; RT 1.45 (Method B)

EXAMPLE 856-[6-(3,4-Dihydroisoquinolin-2(1H)-ylcarbonyl)-1,3-benzodioxol-5-yl]-N-ethyl-N-(4-hydroxyphenyl)-1,2,3,4-tetrahydropyrrolo[1,2-a]pyrazine-8-carboxamide

LC/MS (C₃₃H₃₂N₄O₅) 565 [M+H]⁺; RT 1.49 (Method B)

EXAMPLE 86N-(4-Hydroxyphenyl)-3-[6-[(3R)-3-methyl-3,4-dihydro-1H-isoquinoline-2-carbonyl]-1,3-benzodioxol-5-yl]-N-(3-methylpyrazolo[1,5-a]pyrimidin-6-yl)-indolizine-1-carboxamidehydrochloride Step A:N-[4-[tert-Butyl(dimethyl)silyl]oxyphenyl]-3-[6-[(3R)-3-methyl-3,4-dihydro-1H-isoquinoline-2-carbonyl]-1,3-benzodioxol-5-yl]-N-(3-methylpyrazolo[1,5-a]pyrimidin-6-yl)indolizine-1-carboxamide

To a solution of 0.6 g of3-(6-{[(3R)-3-methyl-3,4-dihydroisoquinolin-2(1H)-yl]carbonyl}-1,3-benzodioxol-5-yl)indolizine-1-carboxylicacid (1.3 mmol) in 6 mL of dichloroethane there is added 0.18 mL of1-chloro-N,N,2-trimethyl-prop-1-en-1-amine (2 mmol). The reactionmixture is stirred at ambient temperature for 2 hours and there is thenadded 0.8 g of the compound of Preparation 36″ (2.2 mmol). The batch isrefluxed for 20 hours and is then cooled and diluted with a mixture ofdichloromethane and saturated NaHCO₃ solution. After separation of thephases, the organic phase is dried over MgSO₄ and concentrated todryness The crude product thereby obtained is purified by chromatographyover silica gel (dichloromethane/methanol gradient).

LC/MS: [M+H]⁺=791.4 vs. 791.3 calculated

Step B:N-(4-Hydroxyphenyl)-3-[6-[(3R)-3-methyl-3,4-dihydro-1H-isoquinoline-2-carbonyl]-1,3-benzodioxol-5-yl]-N-(3-methylpyrazolo[1,5-a]pyrimidin-6-yl)indolizine-1-carboxamidehydrochloride

The procedure is in accordance with a protocol analogous to thatdescribed in Step D of Example 1. The product thereby obtained issubjected to a step of conversion into a salt in the presence of HCl inether.

IR (ATR) cm⁻¹: 2500 to 3000 ν —OH, 1614 ν >C═O amides, 1236 ν >C—O—C<,740 γ >CH—Ar

High-Resolution Mass Spectroscopy (ESI+):

Empirical formula: C₄₀H₃₂N₆O₅

[M+H]⁺, calculated: 677.2507

[M+H]⁺, measured: 677.2506

Pharmacological Study EXAMPLE A Inhibition of Bcl-2 by the FluorescencePolarisation Technique

The fluorescence polarisation tests were carried out on microplates (384wells). The Bcl-2 protein, labelled (histag-Bcl-2 such that Bcl-2corresponds to the UniProtKB® primary accession number: P10415), at afinal concentration of 2.50×10⁻⁸ M, is mixed with a fluorescent peptide(Fluorescein-REIGAQLRRMADDLNAQY), at a final concentration of 1.00×10⁻⁸M in a buffer solution (Hepes 10 mM, NaCl 150 mM, Tween20.05%, pH 7.4),in the presence or absence of increasing concentrations of testcompounds. After incubation for 2 hours, the fluorescence polarisationis measured.

The results are expressed in IC₅₀ (the concentration of compound thatinhibits fluorescence polarisation by 50%) and are presented in Table 1below.

The results show that the compounds of the invention inhibit interactionbetween the Bcl-2 protein and the fluorescent peptide describedhereinbefore.

EXAMPLE B In Vitro Cytotoxicity

The cytotoxicity studies were carried out on the RS4; 11 leukaemiatumour line.

The cells are distributed onto microplates and exposed to the testcompounds for 48 hours.

The cell viability is then quantified by a colorimetric assay, theMicroculture Tetrazolium Assay (Cancer Res., 1987, 47, 939-942).

The results are expressed in IC₅₀ (the concentration of compound thatinhibits cell viability by 50%) and are presented in Table 1 below.

The results show that the compounds of the invention are cytotoxic.

TABLE 1 IC₅₀ of Bcl-2 inhibition (fluorescence polarisation test) and ofcytotoxicity for RS4; 11 cells IC₅₀ (nM) Bcl-2 FP IC₅₀ (nM) MTT RS4; 11Example 1 17.9 11.3 Example 2 17.0 36 Example 3 33.6 66.5 Example 4 56.4251 Example 5 55.9 416 Example 6 60.3 161 Example 7 46.4 108 Example 824.5 20.5 Example 9 40.6 780 Example 10 24.7 439 Example 11 10.9 83.7Example 12 10.4 116 Example 13 5.8 33.65 Example 14 3.7 7.6 Example 155.7 166 Example 16 7.5 252 Example 17 3.4 11.8 Example 18 7.5 47.7Example 19 8.0 235 Example 20 11.1 205 Example 21 4.6 25.3 Example 2212.9 263 Example 23 3.8 9.99 Example 24 6.2 28.4 Example 25 7.9 30Example 26 16.6 300 Example 27 7.7 44.1 Example 28 8.8 112 Example 2919.0 163 Example 30 10.4 52.3 Example 31 5.4 13.7 Example 32 5.0 32.7Example 33 4.6 6.33 Example 34 5.6 27.3 Example 35 15.1 62.2 Example 3612.6 49.7 Example 37 2.9 24.7 Example 38 4.6 9.52 Example 39 4.6 26.3Example 40 6.0 49 Example 41 41.5 294 Example 42 5.1 57.6 Example 43 4.826 Example 44 2.9 8.56 Example 45 3.8 63.8 Example 46 4.1 27.9 Example47 4.3 90.1 Example 48 3.6 24.7 Example 49 3.7 84.7 Example 50 2.2 28.2Example 51 4.8 68.8 Example 52 7.9 20.9 Example 53 5.4 70.9 Example 546.6 45 Example 55 5.5 22.8 Example 56 4.7 36.7 Example 57 21.2 282Example 58 6.4 68.5 Example 59 4.0 21.2 Example 60 5.4 60.3 Example 617.0 61.3 Example 62 5.6 96.6 Example 63 6.2 25.4 Example 64 7.8 282Example 65 5.3 62.8 Example 66 4.7 42 Example 67 ND ND Example 68 8.382.4 Example 69 4.6 1.38 Example 70 5.2 6.17 Example 71 49 ND Example 7290.2 1520 Example 73 83.6 1320 Example 74 68.7 1340 Example 75 67.7 1360Example 76 77.6 1630 Example 77 25.1% @ 10 μM 1880 Example 78 823.3 1880Example 79 99.1 1010 Example 80 299.3 1880 Example 81 12.1 778 Example82   42% @ 10 μM 1880 Example 83 35.8 1500 Example 84 524.9 ND Example85 242.7 ND Example 86 5 20.1 ND: not determined

For partial inhibitors, the percentage fluorescence polarisationinhibition for a given concentration of the test compound is indicated.Accordingly, 25.1% @10 μM means that 25.1% fluorescence polarisationinhibition is observed for a concentration of test compound equal to 10μM.

EXAMPLE C Induction of Caspase Activity In Vivo

The ability of the compounds of the invention to activate caspase 3 isevaluated in a xenograft model of RS4; 11 leukaemia cells.

1×10⁷ RS4;11 cells are grafted sub-cutaneously into immunosuppressedmice (SCID strain). 25 to 30 days after the graft, the animals aretreated orally with the various compounds. Sixteen hours aftertreatment, the tumour masses are recovered and lysed, and the caspase 3activity is measured in the tumour lysates.

This enzymatic measurement is carried out by assaying the appearance ofa fluorigenic cleavage product (DEVDase activity, Promega). It isexpressed in the form of an activation factor corresponding to the ratiobetween the two caspase activities: the activity for the treated micedivided by the activity for the control mice.

The results obtained show that the compounds of the invention arecapable of inducing apoptosis in RS4; 11 tumour cells in vivo.

EXAMPLE D Quantification of the Cleaved Form of Caspase 3 In Vivo

The ability of the compounds of the invention to activate caspase 3 isevaluated in a xenograft model of RS4; 11 leukaemia cells.

1×10⁷ RS4;11 cells are grafted sub-cutaneously into immunosuppressedmice (SCID strain). 25 to 30 days after the graft, the animals aretreated orally with the various compounds. After treatment, the tumourmasses are recovered (after a time period T) and lysed, and the cleaved(activated) form of caspase 3 is quantified in the tumour lysates.

The quantification is carried out using the “Meso Scale Discovery (MSD)ELISA platform” test, which specifically assays the cleaved form ofcaspase 3. It is expressed in the form of an activation factorcorresponding to the ratio between the quantity of cleaved caspase 3 inthe treated mice divided by the quantity of cleaved caspase 3 in thecontrol mice.

The results show that the compounds of the invention are capable ofinducing apoptosis in RS4;11 tumour cells in vivo.

TABLE 2 Caspase activation factors (cleaved caspase 3 MSD test in thetumours of treated mice versus control mice) in vivo, after treatment bythe oral route (exact doses in brackets) Time period after which thetumour Activation factor ± Compound tested is removed (T) SEM (versuscontrol) Example 2 6 hours 14.6 (50 mg/kg) Example 13 2 hours 23.1 (50mg/kg) Example 17 2 hours 15.3 (50 mg/kg) Example 21 2 hours 24.8 ± 1.4(50 mg/kg) Example 32 2 hours 54.4 ± 2.8 (25 mg/kg) Example 33 2 hours31.1 ± 10.8 (25 mg/kg) Example 38 2 hours 27.5 ± 2.6 (25 mg/kg) Example39 2 hours 34.1 ± 2.4 (25 mg/kg) Example 42 2 hours 77.5 ± 4.8 (25mg/kg) Example 50 2 hours 45.2 ± 3.9 (25 mg/kg) Example 56 2 hours 10.3± 4.2 (25 mg/kg)

EXAMPLE E Anti-Tumour Activity In Vivo

The anti-tumour activity of the compounds of the invention is evaluatedin a xenograft model of RS4; 11 leukaemia cells.

1×10⁷ RS4;11 cells are grafted sub-cutaneously into immunosuppressedmice (SCID strain). 25 to 30 days after the graft, when the tumour masshas reached about 150 mm³, the mice are treated orally with the variouscompounds in two different regimes (daily treatment for five days perweek for two weeks, or two treatments weekly for two weeks).

The tumour mass is measured twice weekly from the start of treatment.

The results obtained accordingly show that the compounds of theinvention are capable of inducing significant tumour regression duringthe treatment period.

EXAMPLE F Pharmaceutical Composition: Tablets

1000 tablets containing a dose of 5 mg of a compound 5 g selected fromExamples 1 to 86 Wheat starch 20 g  Maize starch 20 g  Lactose 30 g Magnesium stearate 2 g Silica 1 g Hydroxypropylcellulose 2 g

1-24. (canceled)
 25. A compound of formula (I):

wherein: X and Y represent a carbon atom or a nitrogen atom, it beingunderstood that they may not simultaneously represent two carbons atomsor two nitrogen atoms; the Het moiety of the group

 represents an optionally substituted, aromatic or non-aromatic ringcomposed of 5, 6 or 7 ring members, which may have, in addition to thenitrogen represented by X or by Y, from one to 3 hetero atoms selectedindependently from oxygen, sulphur and nitrogen, it being understoodthat the nitrogen in question may be substituted by a group representinga hydrogen atom, a linear or branched (C₁-C₆)alkyl group or a—C(O)—O-Alk group wherein Alk is a linear or branched (C₁-C₆)alkylgroup; T represents a hydrogen atom, a linear or branched (C₁-C₆)alkylgroup optionally substituted by from one to three halogen atoms, a(C₂-C₄)alkyl-NR₁R₂ group, or a (C₁-C₄)alkyl-OR₆ group; R₁ and R₂independently of one another represent a hydrogen atom or a linear orbranched (C₁-C₆)alkyl group, or R₁ and R₂, together with the nitrogenatom carrying them, form a heterocycloalkyl; R₃ represents a linear orbranched (C₁-C₆)alkyl group, a linear or branched (C₂-C₆)alkenyl group,a linear or branched (C₂-C₆)alkynyl group, a cycloalkyl group, a(C₃-C₁₀)cycloalkyl-(C₁-C₆)alkyl group wherein the alkyl moiety is linearor branched, a heterocycloalkyl group, an aryl group or a heteroarylgroup, it being understood that one or more of the carbon atoms of thepreceding groups, or of their possible substituents, may be deuterated;R₄ represents an aryl group, a heteroaryl group, a cycloalkyl group or alinear or branched (C₁-C₆)alkyl group, it being understood that one ormore of the carbon atoms of the preceding groups, or of their possiblesubstituents, may be deuterated; R₅ represents a hydrogen or halogenatom, a linear or branched (C₁-C₆)alkyl group, or a linear or branched(C₁-C₆)alkoxy group; R₆ represents a hydrogen atom or a linear orbranched (C₁-C₆)alkyl group; R_(a), R_(b), R_(c) and R_(d), eachindependently of the others, represent a hydrogen atom, a linear orbranched (C₁-C₆)alkyl group, a linear or branched (C₂-C₆)alkenyl group,a linear or branched (C₂-C₆)alkynyl group, an aryl group or a heteroarylgroup, a halogen atom, a linear or branched (C₁-C₆)alkoxy group, ahydroxy group, a linear or branched (C₁-C₆)polyhaloalkyl group, atrifluoromethoxy group, —NR₇R₇′, nitro, R₇—CO—(C₀-C₆)alkyl-,R₇—CO—NH—(C₀-C₆)alkyl-, NR₇R₇′—CO—(C₀-C₆)alkyl-,NR₇R₇′—CO—(C₀-C₆)alkyl-O—, R₇—SO₂—NH—(C₀-C₆)alkyl-,R₇—NH—CO—NH—(C₀-C₆)alkyl-, R₇—O—CO—NH—(C₀-C₆)alkyl-, a heterocycloalkylgroup, or the substituents of one of the pairs (R_(a),R_(b)),(R_(b),R_(c)) or (R_(c),R_(d)), together with the carbon atoms carryingthem, form a ring composed of from 5 to 7 ring members, which may havefrom one to 2 hetero atoms selected from oxygen and sulphur, it beingunderstood that one or more carbon atoms of the ring definedhereinbefore may be deuterated or substituted by from one to 3 groupsselected from halogen and linear or branched (C₁-C₆)alkyl; R₇ and R₇′,each independently of the other, represent a hydrogen, a linear orbranched (C₁-C₆)alkyl, a linear or branched (C₂-C₆)alkenyl, a linear orbranched (C₂-C₆)alkynyl, an aryl or a heteroaryl, or R₇ and R₇′,together with the nitrogen atom carrying them, form a heterocyclecomposed of from 5 to 7 ring members; it being understood that: “aryl”means a phenyl, naphthyl, biphenyl or indenyl group, “heteroaryl” meansany mono- or bi-cyclic group composed of from 5 to 10 ring members,having at least one aromatic moiety and containing from 1 to 4 heteroatoms selected from oxygen, sulphur and nitrogen (including quaternarynitrogens), “cycloalkyl” means any mono- or hi-cyclic, non-aromatic,carbocyclic group containing from 3 to 10 ring members,“heterocycloalkyl” means any mono- or bi-cyclic, non-aromatic, condensedor spiro group containing from 3 to 10 ring members and containing from1 to 3 hetero atoms selected from oxygen, sulphur, SO, SO₂ and nitrogen,wherein the aryl, heteroaryl, cycloalkyl and heterocycloalkyl groups sodefined and the groups alkyl, alkenyl, alkynyl and alkoxy may beoptionally substituted by from 1 to 3 groups selected from optionallysubstituted, linear or branched (C₁-C₆)alkyl, (C₃-C₆)spiro, optionallysubstituted, linear or branched (C₁-C₆)alkoxy, (C₁-C₆)alkyl-S—, hydroxy,oxo (or N-oxide where appropriate), nitro, cyano, —COOR′, —OCOR′, NR′R″,linear or branched (C₁-C₆)polyhaloalkyl, trifluoromethoxy,(C₁-C₆)alkylsulphonyl, halogen, optionally substituted aryl, heteroaryl,aryloxy, arylthio, cycloalkyl, heterocycloalkyl optionally substitutedby one or more halogen atoms or alkyl groups, it being understood thatR′ and R″, each independently of the other, represent a hydrogen atom oran optionally substituted, linear or branched (C₁-C₆)alkyl group, andwherein the Het moiety of the group

defined in formula (I) may be optionally substituted by from one tothree groups selected from linear or branched (C₁-C₆)alkyl, hydroxy,linear or branched (C₁-C₆)alkoxy, NR₁′R₁″ and halogen, wherein R₁′ andR₁″, each independently of the other, represent a hydrogen atom or anoptionally substituted, linear or branched (C₁-C₆)alkyl group, itsenantiomers and diastereoisomers, and addition salts thereof with apharmaceutically acceptable acid or base.
 26. The compound according toclaim 25, wherein the group

represents: 5,6,7,8-tetrahydroindolizine optionally substituted by anamino group; indolizine; 1,2,3,4-tetrahydropyrrolo[1,2-a]pyrazineoptionally substituted by a methyl; or pyrrolo[1,2-a]pyrimidine.
 27. Thecompound according to claim 25, wherein T represents hydrogen, methyl,2-(morpholin-4-yl)ethyl, 3-(morpholin-4-yl)propyl, —CH₂—OH,2-aminoethyl, 2-(3,3-difluoropiperidin-1-yl)ethyl,2-[(2,2-difluoroethyl)amino]ethyl or 2-(3-methoxyazetidin-1-yl)ethyl.28. The compound according to claim 25, wherein R_(a) and R_(d) eachrepresent a hydrogen atom and (R_(b),R_(c)), together with the carbonatoms carrying them, form a 1,3-dioxolane group or a 1,4-dioxane group;or R_(a), R_(c) and R_(d) each represent a hydrogen atom and R_(b)represents a hydrogen, a halogen, a methyl or a methoxy; or R_(a), R_(b)and R_(d) each represent a hydrogen atom and R_(c) represents a hydroxyor methoxy group.
 29. The compound according to claim 25, wherein R₄represents a 4-hydroxyphenyl group.
 30. The compound according to claim25, wherein R₃ represents a linear (C₁-C₆)alkyl, aryl or heteroarylgroup, wherein the latter two groups may be optionally substituted byfrom one to three groups selected from halogen, linear or branched(C₁-C₆)alkyl, linear or branched (C₁-C₆)alkoxy and cyano.
 31. Thecompound according to claim 25, wherein R₃ represents a heteroaryl groupselected from: 1H-indole, 2,3-dihydro-1H-indole, 1H-indazole, pyridine,1H-pyrrolo[2,3-b]pyridine, 1H-pyrazole, imidazo[1,2-a]pyridine,pyrazolo[1,5-a]pyrimidine, [1,2,4]triazolo[1,5-a]pyrimidine, and1H-pyrazolo[3,4-b]pyridine, all of which may be optionally substitutedby a linear or branched (C₁-C₆)alkyl group.
 32. The compound accordingto claim 25, which is selected from:N-(4-hydroxyphenyl)-3-(6-{[(3R)-3-methyl-3,4-dihydroisoquinolin-2(IH)-yl]-carbonyl)}-1,3-benzodioxol-5-yl)-N-{1-[2-(morpholin-4-yl)ethyl]-1H-indol-5-yl}-5,6,7,8-tetrahydroindolizine-1-carboxamide,N-(4-hydroxyphenyl)-3-(6-{[(3S)-3-[2-(morpholin-4-yl)ethyl]-3,4-dihydroisoquinolin-2(1H)-yl]carbonyl}-1,3-benzodioxol-5-yl)-N-phenyl-5,6,7,8-tetrahydroindolizine-1-carboxamide,N-{3-fluoro-4-[2-(morpholin-4-yl)ethoxy]phenyl}-N-(4-hydroxyphenyl)-3-(6-{[(3R)-3-methyl-3,4-dihydroisoquinolin-2(1H)-yl]carbonyl}-1,3-benzodioxol-5-yl)indolizine-1-carboxamide,N-(4-hydroxyphenyl)-3-(6-{[(3R)-3-methyl-3,4-dihydroisoquinolin-2(1H)-yl]carbonyl}-1,3-benzodioxol-5-yl)-N-(pyridin-4-yl)indolizine-1-carboxamide,N-(4-hydroxyphenyl)-3-(6-{[(3R)-3-methyl-3,4-dihydroisoquinolin-2(1H)-yl]-carbonyl}-1,3-benzodioxol-5-yl)-N-(2-methylpyridin-4-yl)indolizine-1-carboxamide,N-(4-hydroxyphenyl)-3-(6-{[(3R)-3-methyl-3,4-dihydroisoquinolin-2(1H)-yl]-carbonyl}-1,3-benzodioxol-5-yl)-N-(1-methyl-1H-pyrrolo[2,3-b]pyridin-5-yl)-indolizine-1-carboxamide,N-(4-hydroxyphenyl)-3-(6-{[(3R)-3-[3-(morpholin-4-yl)propyl]-3,4-dihydroisoquinolin-2(1H)-yl]carbonyl}-1,3-benzodioxol-5-yl)-N-phenyl-5,6,7,8-tetrahydroindolizine-1-carboxamide,N-(2,6-dimethylpyridin-4-yl)-N-(4-hydroxyphenyl)-3-(6-{[(3R)-3-methyl-3,4-dihydroisoquinolin-2(H)-yl]carbonyl}-1,3-benzodioxol-5-yl)indolizine-1-carboxamide,N-(4-hydroxyphenyl)-3-(6-{[(3R)-3-methyl-3,4-dihydroisoquinolin-2(1H)-yl]-carbonyl}-1,3-benzodioxol-5-yl)-N-(pyridin-4-yl)-5,6,7,8-tetrahydroindolizine-1-carboxamide,3-(5-chloro-2-{[(3R)-3-methyl-3,4-dihydroisoquinolin-2(1H)-yl]carbonyl}phenyl)-N-(4-hydroxyphenyl)-N-(1-methyl-1H-pyrrolo[2,3-b]pyridin-5-yl)indolizine-1-carboxamide,N-(4-hydroxyphenyl)-N-(2-methoxypyridin-4-yl)-3-(6-{[(3R)-3-methyl-3,4-dihydroisoquinolin-2(1H)-yl]carbonyl}-1,3-benzodioxol-5-yl)indolizine-1-carboxamide,and their enantiomers and diastereoisomers, and addition salts thereofwith a pharmaceutically acceptable acid or base.
 33. A pharmaceuticalcomposition comprising the compound according to claim 25, incombination with one or more pharmaceutically acceptable excipients. 34.A method of treating a condition requiring a pro-apoptotic agent in asubject in need thereof, comprising administration of a compoundaccording to claim 25, alone or in combination with one or morepharmaceutically acceptable excipients.
 35. The method according toclaim 34, wherein the condition is selected from cancers, auto-immunediseases and diseases of the immune system.
 36. The method according toclaim 34, wherein the condition is selected from cancers of the bladder,brain, breast and uterus, chronic lymphoid leukaemias, colorectalcancer, cancers of the œsophagus and liver, lymphoblastic leukaemias,non-Hodgkin lymphomas, melanomas, malignant haemopathies, myelomas,ovarian cancer, non-small-cell lung cancer, prostate cancer andsmall-cell lung cancer.
 37. A combination of a compound according toclaim 25 with an anti-cancer agent selected from genotoxic agents,mitotic poisons, anti-metabolites, proteasome inhibitors, kinaseinhibitors and antibodies.
 38. A pharmaceutical composition comprising acombination according to claim 37, in combination with one or morepharmaceutically acceptable excipients.
 39. A method of treating cancerin a subject in need thereof, comprising administration of a combinationaccording to claim
 37. 40. A method of treating cancer in a subject inneed thereof comprising administration of a compound according to claim25 in combination with radiotherapy.